Chromane compounds

ABSTRACT

The present invention provides a compound which is useful as an active ingredient of a pharmaceutical composition, in particular, a pharmaceutical composition for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including a pharmaceutical composition for preventing or treating diseases including, but not limited to, Glaucoma, MCI (Mild cognitive impairment) or Alzheimer&#39;s disease, especially, Alzheimer&#39;s disease.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of International Application No.PCT/US2013/043016 having an international filing date of May 29, 2013,which claims priority benefit of U.S. Provisional Patent ApplicationNos. 61/653,321 filed on May 30, 2012 and 61/782,038 filed on Mar. 14,2013, the disclosures of which are incorporated herein by reference intheir entirety.

INCORPORATION BY REFERENCE

The content of the following submission on ASCII text file (ST.25 textformat) is incorporated herein by reference in its entirety: a computerreadable form (CRF) of the Sequence Listing (file name is“322732001040_Sequence_Listing.txt”; date recorded: May 9, 2013; and thesize of the ASCII text file in bytes is 4,096 bytes).

TECHNICAL FIELD

The present invention relates to a chromane compound which is useful asan active ingredient of a pharmaceutical composition, in particular, apharmaceutical composition for preventing or treating diseases orconditions associated with and/or mediated by β-secretase activity,hydrolysis of a β-secretase site of an amyloid precursor protein, and/orβ-amyloid protein accumulation, including a pharmaceutical compositionfor preventing or treating diseases including, but not limited to,Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease,especially, Alzheimer's disease.

BACKGROUND OF THE INVENTION

Alzheimer's disease is a progressive mental deterioration in a humanresulting, inter alia, in loss of memory, confusion and disorientation.Alzheimer's disease accounts for the majority of senile dementia and isa leading cause of death in adults (Non-Patent Document 1).Histologically, the brain of persons afflicted with Alzheimer's diseaseis characterized by a distortion of the intracellular neurofibrils andthe presence of senile plaques composed of granular or filamentousargentophilic masses with an amyloid protein core, largely due to theaccumulation of β-amyloid protein (Aβ) in the brain. Aβ accumulationplays a role in the pathogenesis and progression of the disease(Non-Patent Document 2) and is a proteolytic fragment of amyloidprecursor protein (APP). APP is cleaved initially by β-secretasefollowed by γ-secretase to generate Aβ (Non-Patent Document 3 and 4).

It is known that inhibition of BACE may have a therapeutic effect in theprevention of dementia after stroke recovery (Non-Patent Document 5). Itis reported that inhibition of BACE1 (beta-secretase 1) may have atherapeutic effect in Down syndrome (Non-Patent Document 6). Therelationship between BACE1 mRNA levels and Parkinson's disease (PD) andDementia with Lewy bodies (DLB) is also reported (Non-Patent Document 7and 8).

In Patent Document 1, it is described that compounds (A) which are BACEinhibitors and are useful as therapeutic agents in the treatment,prevention, and amelioration of a disease or disorder characterized byelevated β-amyloid deposits or β-amyloid levels in a patient.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 2, it is described that compounds (B) which areuseful for inhibition of β-secretase enzymatic activity and for therapyand/or prophylaxis of neurodegenerative diseases associated therewith,particularly Alzheimer's Disease.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 3, it is described that compounds (C) which are BACEinhibitors and are useful as therapeutic agents in the treatment,prevention, and amelioration of a disease or disorder characterized byelevated β-amyloid deposits or β-amyloid levels in a patient.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 4, it is described that compounds (D) which are BACEinhibitors and are useful as therapeutic agents in the treatment,prevention, and amelioration of a disease or disorder characterized byelevated β-amyloid deposits or β-amyloid levels in a patient.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 5 and 6, it is described that compounds (E) haveBACE1 inhibitory activity and are useful as prophylactic or therapeuticagent for a neurodegenerative disease caused by Aβ and typified byAlzheimer type dementia, and pharmaceutical use thereof.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 7, it is described that compounds (F) are useful forthe modulation of the beta-secretase activity and are useful for thetreatment of Alzheimer's disease and beta-secretase and/or plaquemediated disorders.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 8, it is described that compounds (G) are useful forthe modulation of the beta-secretase activity and are useful for thetreatment of Alzheimer's disease and beta-secretase and/or plaquemediated disorders.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 9, it is described that compounds (H) are useful forthe modulation of the beta-secretase activity and are useful for thetreatment of Alzheimer's disease and beta-secretase and/or plaquemediated disorders.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 10, it is described that compounds (I) are useful forinhibition of β-secretase enzyme activity and the therapy and/orprophylaxis of neurodegenerative diseases associated therewith, such asAlzheimer's disease.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 11, it is described that compounds (J) are inhibitorsof beta-secretase-2 (BACE2) and the compounds may therefore be useful inthe treatment of type 2 diabetes and other metabolic disorders.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 12, it is described that compounds (K) are useful forinhibition of β-secretase enzyme activity and the therapy and/orprophylaxis of neurodegenerative diseases associated therewith, such asAlzheimer's disease.

(for the symbols in the formula, refer to the patent publication).

In Patent Document 13, it is described that compounds (L) are inhibitorsof β-secretase and hence inhibit the formation of amyloid β (Aβ)peptides and are useful for treatment and/or prevention of Aβ-relatedpathologies such as Alzheimer's disease, and so on.

(for the symbols in the formula, refer to the patent publication).

In any of these Patent Documents, there is no specific disclosure of thecompound of the present invention.

REFERENCES

-   Patent Document 1: Pamphlet of International Publication WO    2010/021680-   Patent Document 2: Pamphlet of International Publication WO    2011/072064-   Patent Document 3: Pamphlet of International Publication WO    2011/106414-   Patent Document 4: Pamphlet of International Publication WO    2010/105179-   Patent Document 5: Pamphlet of International Publication WO    2010/013302-   Patent Document 6: Pamphlet of International Publication WO    2010/013794-   Patent Document 7: Pamphlet of International Publication WO    2010/030954-   Patent Document 8: Pamphlet of International Publication WO    2011/115938-   Patent Document 9: Pamphlet of International Publication WO    2011/115928-   Patent Document 10: Pamphlet of International Publication    WO2011/123674-   Patent Document 11: Pamphlet of International Publication    WO2010/128058-   Patent Document 12: Pamphlet of International Publication    WO2012/071458-   Patent Document 13: Pamphlet of International Publication    WO2012/087237-   Non-Patent Document 1: Anderson, R. N., et al., Natl. Vital Stat.    Rep. 49:1-87 (2001)-   Non-Patent Document 2: Selkoe, D. J., Nature 399: 23-31 (1999)-   Non-Patent Document 3: Lin, X., et al., Proc. Natl. Acad. Sci. USA    97:1456-1460 (2000)-   Non-Patent Document 4: De Stropper, B., et al., Nature 391:387-390    (1998)-   Non-Patent Document 5: Wen Y., et al., Brain Res. 1009 (1-2):1-8    (2004)-   Non-Patent Document 6: Miners J. S., et al., J. Alzheimer's Dis. 23    (1):101-108 (2011)-   Non-Patent Document 7: Coulson D T., et al., J. Alzheimer's Dis. 22    (4):1111-1122 (2010)-   Non-Patent Document 8: Halliday G M., et al., J. Neural Transm. 118    (5):713-719 (2011)

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 Drawing shows VCD spectra of Ex. 228b compound.

FIG. 2 Drawing shows VCD spectra of Ex. 229b compound.

FIG. 3 Drawing shows VCD spectra of Reference Example 225a compound.

SUMMARY OF THE INVENTION

The present invention provides a compound which is useful as an activeingredient of a pharmaceutical composition, in particular, apharmaceutical composition for preventing or treating diseases orconditions associated with and/or mediated by β-secretase activity,hydrolysis of a β-secretase site of a β-amyloid precursor protein,and/or β-amyloid protein accumulation, including a pharmaceuticalcomposition for preventing or treating diseases including, but notlimited to, Glaucoma, MCI (Mild cognitive impairment) or Alzheimer'sdisease, especially, Alzheimer's disease.

Means for Solving the Problems

The present inventors have extensively studied compounds havingbeta-secretase inhibitory activity, and as a result, they have foundthat chromane compounds which are the compounds of the present inventionhave excellent beta-secretase inhibitory activity, and are thereforeuseful as agents for preventing or treating diseases or conditionsassociated with and/or mediated by β-secretase activity, hydrolysis of aβ-secretase site of a β-amyloid precursor protein, and/or β-amyloidprotein accumulation, including a pharmaceutical composition forpreventing or treating diseases including, but not limited to, Glaucoma,MCI (Mild cognitive impairment) or Alzheimer's disease, especially,Alzheimer's disease, thereby completing the present invention.

The present invention relates to compounds of the formula (I) or a saltthereof:

wherein

A¹ is O, S, —C(R^(A11)R^(A12)) T-, or -T-C(R^(A11)R^(A12))—;

A² is —C(R^(A21)R^(A22))—;

T is a single bond, O, or S;

R^(A11), R^(A12), R^(A21) and R^(A22) are, independently, H or halogen;or

R^(A11), R^(A12), R^(A21) and R^(A22) are combined with each other toform an aryl group, which is unsubstituted or substituted;

B is a hetero ring group which is unsubstituted or substituted, orcycloalkyl which is unsubstituted or substituted;

X and Y are independently selected from the group consisting of

H, lower alkyl, which is unsubstituted or substituted, and cycloalkyl,which is unsubstituted or substituted; or

X and Y are combined with each other to form a cycloalkyl group, whichis unsubstituted or substituted; and

R¹, R², R³ and R⁴ are independently selected from the group consistingof

H, halogen, lower alkyl, which is unsubstituted or substituted, loweralkenyl, which is unsubstituted or substituted, —N(H)-(hetero ringgroup), wherein said hetero ring group is unsubstituted or substituted,—N(H)—C(O)-(hetero ring group), wherein said hetero ring group isunsubstituted or substituted, cycloalkenyl, which is unsubstituted orsubstituted, aryl, which is unsubstituted or substituted, and a heteroring group, which is unsubstituted or substituted.

Further, unless specifically described otherwise, in the case where thesymbols in any of the formulae in the present specification are alsoused in other formulae, the same symbols denote the same meanings.

Furthermore, the present invention relates to pharmaceuticalcompositions, comprising compounds of formula (I) or a salt thereof, asdescribed herein, and a pharmaceutically acceptable carrier. Moreover,the present invention relates to pharmaceutical compositions forpreventing or treating diseases or conditions associated with and/ormediated by β-secretase activity, hydrolysis of a β-secretase site of aβ-amyloid precursor protein, and/or β-amyloid protein accumulation,including compounds of formula (I) or a salt thereof, as describedherein, that are agents for preventing or treating diseases orconditions associated with and/or mediated by β-secretase activity,hydrolysis of a β-secretase site of a β-amyloid precursor protein,and/or β-amyloid protein accumulation, including compounds of formula(I) or a salt thereof.

Furthermore, the present invention relates to use of compounds offormula (I) or a salt thereof, as described herein, for preparation of apharmaceutical composition (e.g., medicament) for preventing or treatingdiseases or conditions associated with and/or mediated by β-secretaseactivity, hydrolysis of a β-secretase site of a β-amyloid precursorprotein, and/or β-amyloid protein accumulation, use of compounds offormula (I) or a salt thereof for preventing or treating diseases orconditions associated with and/or mediated by β-secretase activity,hydrolysis of a β-secretase site of a β-amyloid precursor protein,and/or β-amyloid protein accumulation, and methods for preventing ortreating diseases or conditions associated with and/or mediated byβ-secretase activity, hydrolysis of a β-secretase site of a β-amyloidprecursor protein, and/or β-amyloid protein accumulation, includingadministering to a subject in need thereof an effective amount of thecompounds of formula (I) or a salt thereof.

The present invention also relates to compounds of formula (I) or a saltthereof, as described herein, for use in the prevention or treatment ofdiseases or conditions associated with and/or mediated by β-secretaseactivity, hydrolysis of a β-secretase site of a β-amyloid precursorprotein, and/or β-amyloid protein accumulation, the compounds of formula(I) or a salt thereof for preventing or treating diseases or conditionsassociated with and/or mediated by β-secretase activity, hydrolysis of aβ-secretase site of a β-amyloid precursor protein, and/or β-amyloidprotein accumulation. The present invention also relates to a method forpreventing or treating diseases or conditions associated with and/ormediated by β-secretase activity, hydrolysis of a β-secretase site of aβ-amyloid precursor protein, and/or β-amyloid protein accumulation,including administering to a subject an effective amount of thecompounds of formula (I) or a salt thereof.

Effects of the Invention

The compounds of formula (I) or a salt thereof have beta-secretaseinhibitory activity, and therefore can be used as an agent forpreventing or treating diseases or conditions associated with and/ormediated by β-secretase activity, hydrolysis of a β-secretase site of aβ-amyloid precursor protein, and/or β-amyloid protein accumulation,including, but not limited to, diseases such as Glaucoma, MCI (Mildcognitive impairment) or Alzheimer's disease, especially, Alzheimer'sdisease, or the like. In some embodiments, the compounds of formula (I)or a salt thereof can be used as an agent for preventing or treatingdiseases or conditions including, but not limited to, stroke,cerebrovascular dementia, Down syndrome, Parkinson's disease (PD), anddementia with Lewy bodies (DLB).

DETAILED DESCRIPTION

The present invention will be explained in more detail herein below.Further, “the compounds of formula (I) or a salt thereof” may be denotedas “the compounds (I) of the present invention” or “the compounds (I)”below in some cases.

In the present specification, the term “lower alkyl” refers to astraight (linear) or branched chain alkyl having 1 to 6 carbon atoms(hereinafter simply referred to as C₁₋₆), for example, methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl,n-hexyl, and the like. In another embodiment, it is C₁₋₄ alkyl, and in afurther embodiment, C₁₋₃ alkyl.

The term “lower alkenyl” refers to a straight (linear) or branched chainC₂₋₆ alkenyl, for example, vinyl, propenyl, butenyl, pentenyl,1-methylvinyl, 1-methyl-2-propenyl, 1,3-butadienyl, 1,3-pentadienyl, orthe like. In another embodiment, it is C₂₋₄ alkenyl, and in a stillanother embodiment, C₂₋₃ alkenyl.

The term “lower alkynyl” refers to a linear or branched chain C₂₋₆alkynyl, for example, ethynyl, propynyl, butynyl, pentynyl,1-methyl-2-propynyl, 1,3-butadiynyl, 1,3-pentadiynyl, or the like. Inanother embodiment, it is C₂₋₄ alkynyl.

The term “cycloalkyl” refers to a C₃₋₁₀ saturated hydrocarbon ringgroup, which may have a bridge. It is, for example, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl,or the like, in another embodiment, C₃₋₈ cycloalkyl, and in a furtherembodiment, C₃₋₆ cycloalkyl.

The term “cycloalkenyl” refers to a C₄₋₁₅ hydrocarbon ring group havingat least one double bond in the ring (provided that an aromatichydrocarbon ring group is excluded), which may have a bridge, andincludes a ring group fused (e.g., condensed) with a benzene ring at adouble bond site. It is, for example, cyclopentenyl, cyclopentadienyl,cyclohexenyl, cyclohexadienyl, 1-tetrahydronaphthyl, 1-indenyl,9-fluorenyl, or the like. In another embodiment, it is C₅₋₁₀cycloalkenyl, in a further embodiment, C₅₋₈ cycloalkenyl, and in afurther embodiment, C₅₋₇ cycloalkenyl.

The term “aryl” refers to a C₆₋₁₄ monocyclic to tricyclic aromatichydrocarbon ring group, and includes a ring group fused with C₅₋₈cycloalkene at its double bond site. It is, for example, phenyl,naphthyl, 5-tetrahydronaphthyl, 4-indenyl, 1-fluorenyl, or the like. Andthe term “aryl” does not encompass aryl rings containing hetero atoms(such as S, N, O).

The term “hetero ring” means a ring group containing i) a monocyclic 3-to 8-membered hetero ring containing 1 to 4 hetero atoms selected fromoxygen, sulfur, and nitrogen, and in another embodiment, a 5- to7-membered hetero ring containing 1 to 4 hetero atoms selected fromoxygen, sulfur, and nitrogen, and ii) a bicyclic or tricyclic heteroring (in which the bicyclic or tricyclic heterocyclic ring may include aspiro ring) containing 1 to 5 hetero atoms selected from oxygen, sulfur,and nitrogen, formed by condensation or ring-fusion of the monocyclichetero ring with one or two rings selected from the group consisting ofa monocyclic hetero ring, a benzene ring, C₅₋₈ cycloalkane, and C₅₋₈cycloalkene. The ring atom, sulfur or nitrogen, may be oxidized to forman oxide or a dioxide.

Examples of the “hetero ring” group include the following embodiments:

(1) Monocyclic Saturated Hetero Ring Groups, which mean monocyclic 3- to8-membered saturated rings containing 1 to 4 hetero atoms selected fromoxygen, sulfur, and nitrogen, and in another embodiment, 5- to7-membered hetero rings containing 1 to 4 hetero atoms selected fromoxygen, sulfur, and nitrogen.

(a) those containing 1 to 4 nitrogen atoms, for example, azepanyl,diazepanyl, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl,piperidyl, piperazolidinyl, piperazinyl, azocanyl, hexamethyleneimino,homopiperazinyl, and the like;

(b) those containing 1 to 3 nitrogen atoms and 1 to 2 sulfur atomsand/or 1 to 2 oxygen atoms, for example, thiomorpholinyl, thiazolidinyl,isothiazolidinyl, oxazolidinyl, morpholinyl, and the like;

(c) those containing 1 to 2 sulfur atoms, for example,tetrahydrothiopyranyl and the like;

(d) those containing 1 to 2 sulfur atoms and 1 to 2 oxygen atoms, forexample, oxathiolanyl and the like; and

(e) those containing 1 to 2 oxygen atoms, for example, oxiranyl,oxetanyl, dioxolanyl, tetrahydrofuranyl, tetrahydropyranyl,1,4-dioxanyl, and the like;

(2) Monocyclic Unsaturated Hetero Ring Groups, which mean monocyclic 3-to 8-membered unsaturated rings containing 1 to 4 hetero atoms selectedfrom oxygen, sulfur, and nitrogen, and in another embodiment, 5- to7-membered hetero rings containing 1 to 4 hetero atoms selected fromoxygen, sulfur, and nitrogen.

(a) those containing 1 to 4 nitrogen atoms, for example, pyrrolyl,2-pyrrolinyl, imidazolyl, 2-imidazolinyl, pyrazolyl, 2-pyrazolinyl,pyridyl, dihydropyridyl, tetrahydropyridinyl, pyrimidinyl, pyrazinyl,pyridazinyl, triazolyl, tetrazolyl, triazinyl, dihydrotriazinyl,azepinyl, and the like;

(b) those containing 1 to 3 nitrogen atoms and 1 to 2 sulfur atomsand/or 1 to 2 oxygen atoms, for example, thiazolyl, isothiazolyl,thiadiazolyl, dihydrothiazinyl, oxazolyl, isoxazolyl, oxadiazolyl,oxazinyl, and the like;

(c) those containing 1 to 2 sulfur atoms, for example, thienyl,thiepinyl, dihydrodithiopyranyl, dihydrodithionyl, 2H-thiopyranyl, andthe like;

(d) those containing 1 to 2 sulfur atoms and 1 to 2 oxygen atoms, forexample, dihydroxythiopyranyl and the like; and

(e) those containing 1 to 2 oxygen atoms, for example, furyl,dihydrofuryl, pyranyl, 2H-pyranyl, oxepinyl, dioxolyl, and the like;

(3) Fused Polycyclic Saturated Hetero Ring Groups, which mean bicyclicor tricyclic saturated hetero rings (in which the bicyclic or tricyclicheterocyclic ring may include a spiro ring) containing 1 to 5 heteroatoms selected from oxygen, sulfur, and nitrogen, formed by condensationor ring-fusion of the monocyclic saturated hetero ring with one or tworings selected from the group consisting of a monocyclic saturatedhetero ring, and C₅₋₈ cycloalkane.

(a) those containing 1 to 5 nitrogen atoms, for example, quinuclidinyl,7-azabicyclo[2.2.1]heptyl, 3-azabicyclo[3.2.2]nonanyl,2,8-diazaspiro[4.5]decan-8-yl, 2,3,6,8-tetraazaspiro[4.5]decan-8-yl, andthe like;

(b) those containing 1 to 4 nitrogen atoms and 1 to 3 sulfur atoms,and/or 1 to 3 oxygen atoms, for example, trithiadiazaindenyl,dioxoloimidazolidinyl, 6-oxa-2,8-diazaspiro[4.5]decan-8-yl,6-thia-2,8-diazaspiro[4.5]decan-8-yl, and the like; and

(c) those containing 1 to 3 sulfur atoms and/or 1 to 3 oxygen atoms, forexample, 2,6-dioxabicyclo[3.2.2]oct-7-yl,2-oxa-6-thiaspiro[4.5]decan-8-yl, and the like;

(4) Fused Polycyclic Unsaturated Hetero Ring Groups, which mean bicyclicor tricyclic unsaturated hetero rings (in which the bicyclic ortricyclic heterocyclic ring may include a spiro ring) containing 1 to 5hetero atoms selected from oxygen, sulfur, and nitrogen, formed bycondensation or ring-fusion of the monocyclic hetero ring with one ortwo rings selected from the group consisting of a monocyclic heteroring, a benzene ring, C₅₋₈ cycloalkane, and C₅₋₈ cycloalkene.

(a) those containing 1 to 5 nitrogen atoms, for example, indolyl,isoindolyl, indolinyl, indolizinyl, benzimidazolyl,dihydrobenzimidazolyl, tetrahydrobenzimidazolyl, quinolyl,tetrahydroquinolyl, isoquinolyl, tetrahydroisoquinolyl, indazolyl,imidazopyridyl, benzotriazolyl, tetrazolopyridazinyl, carbazolyl,acridinyl, quinoxalinyl, dihydroquinoxalinyl, tetrahydroquinoxalinyl,phthalazinyl, dihydroindazolyl, benzopyrimidinyl, naphthyridinyl,quinazolinyl, cinnolinyl, pyridopyrrolidinyl, triazolopiperidinyl,9,10-dihydroacridinyl, 2,8-diazaspiro[4.5]deca-3-en-8-yl,2,3,6,8-tetraazaspiro[4.5]deca-1-en-8-yl, and the like;

(b) those containing 1 to 4 nitrogen atoms, and 1 to 3 sulfur atomsand/or 1 to 3 oxygen atoms, for example, benzothiazolyl,dihydrobenzothiazolyl, benzothiadiazolyl, imidazothiazolyl,imidazothiadiazolyl, benzoxazolyl, dihydrobenzoxazolyl,dihydrobenzoxazinyl, benzoxadiazolyl, benzoisothiazolyl,benzoisoxazolyl, thiazolopiperidinyl, 10H-phenothiazinyl,6-oxa-2,8-diazaspiro[4.5]deca-3-en-8-yl,6-thia-2,8-diazaspiro[4.5]deca-3-en-8-yl, and the like;

(c) those containing 1 to 3 sulfur atoms, for example, benzothienyl,benzodithiopyranyl, dibenzo[b,d]thienyl, and the like;

(d) those containing 1 to 3 sulfur atoms and 1 to 3 oxygen atoms, forexample, benzoxathiopyranyl, 2-oxa-6-thiaspiro[4.5]deca-3-en-8-yl, andthe like; and

(e) those containing 1 to 3 oxygen atoms, for example, benzodioxolyl,benzofuranyl, dihydrobenzofuranyl, isobenzofuranyl, chromanyl,chromenyl, isochromenyl, dibenzo[b,d]furanyl, methylenedioxyphenyl,ethylenedioxyphenyl, xanthenyl, and the like; etc.

Further, the terms “aryl”, “cycloalkyl”, and “hetero ring” groups asdescribed above are meant to be monovalent groups, but these may bedivalent or higher groups in some cases. For example, when aryl in theR² is substituted, this aryl is described by monovalent group, but thisaryl means divalent or higher groups.

The term “nitrogen-containing hetero ring” group refers to onecontaining at least one nitrogen atom, including, but not limited to,such as groups in (1)(a), (1)(b), (2)(a), (2)(b), (3)(a), (3)(b),(4)(a), and (4)(b), among the “hetero ring” groups above.

The term “oxygen-containing monocyclic saturated hetero ring” grouprefers to one containing at least one oxygen atom, including, but notlimited to, such as groups containing at least one oxygen atom in (1)(b)or such as groups in (1)(d), and (1)(e), among the “(1) MonocyclicSaturated Hetero Ring Groups” above.

The term “cyclic ether” group refers to one containing only at least oneoxygen atom as hetero atom, including, but not limited to, such asgroups in, (1)(e), among the “oxygen-containing monocyclic saturatedhetero ring” group above.

The term “nitrogen-containing monocyclic hetero ring” group refers toone containing at least one nitrogen atom, including, but not limitedto, such as groups in (1)(a), (1)(b), (2)(a), and (2)(b), among the“Monocyclic Saturated Hetero Ring Groups” and “Monocyclic UnsaturatedHetero Ring Groups” above.

The term “halogen” means F, Cl, Br, or I.

In the present specification, the term “substituted” represents beingsubstituted with 1 to 5 substituents. In some embodiments, the term“substituted” represents being substituted with 1, 2, 3, 4 or 5substituents. Further, if a plurality of substituents are included, thesubstituents may be the same as or different from one another.

In some embodiments, the term “substituted with one or moresubstituents” represents being substituted with 1 to 5 substituents. Insome embodiments, the term “substituted with one or more substituents”represents being substituted with 1, 2, 3, 4 or 5 substituents.

In the present specification, both B and

represent a group that shares a carbon atom with the chromane ring towhich it is attached, as shown in formula (I). For example, when B isoxetanyl, this means that

and when B is cyclopropyl, this means that

In some embodiments, the B group may be substituted at one or moreavailable positions, as described herein.

“R^(A11), R^(A12), R^(A21) and R^(A22) are combined with each other toform an aryl group” indicates that R^(A11), R^(A12), R^(A21) and R^(A22)combined with each carbon atom to which they are bonded to form a C₆₋₁₄monocyclic to tricyclic aromatic hydrocarbon ring group, and includes aring group fused with C₅₋₈ cycloalkene at its double bond site. It is,for example, phenyl, naphthyl, 5-tetrahydronaphthyl, 4-indenyl,1-fluorenyl, or the like.

For example, when R^(A11), R^(A12), R^(A21) and R^(A22) are combinedwith each other to form phenyl, a structure of the compound of formula(I) is as below.

“X and Y are combined with each other to form a cycloalkyl group”indicates that X and Y combined with a carbon atom to which they arebonded to form a C₃₋₁₀ saturated hydrocarbon ring group, which may havea bridge. It is, for example, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, or the like, in anotherembodiment, C₃₋₈ cycloalkyl, and in a further embodiment, C₃₋₆cycloalkyl.

For example, when X and Y are combined with each other to formcyclobutyl or cyclopentyl structures of the compound of formula (I) areas below.

“Amyloid precursor protein,” or “APP,” as used herein, refers to anamyloid precursor polypeptide comprising a β-secretase cleavage site.

A “β-secretase cleavage site” is an amino acid sequence that is cleavedby an active memapsin 2 (also referred to as beta-secretase 1 or BACE-1,or active fragment thereof, such as described in U.S. Pat. No.6,545,127). Specific β-secretase cleavage sites have also beenpreviously set forth and discussed in detail in U.S. Application No.20040121947, and International Application No. PCT/US02/34324(Publication No. WO 03/039454), which are herein incorporated byreference for all purposes in their entirety, and include the Swedishmutation sequence, and the native amyloid precursor protein cleavagesequence. Thus, β-secretase inhibitors may be tested for their abilityto decrease the hydrolysis of the β-secretase cleavage site of asubstrate, such as the amyloid precursor protein, compounds of amyloidprecursor protein, or fragments of amyloid precursor protein.

A “beta-secretase inhibitor” (i.e. β-secretase inhibitor) refers to acompound capable of reducing the proteolytic activity of memapsin-2relative to the activity in the absence of inhibitor.

“Memapsin-2,” as used herein, refers to proteins identified by NationalCenter for Biotechnology Information (“NCBI”) accession numberNP_(—)036236 (sometimes referred to as “β-site APP-cleaving enzyme 1” or“BACE1” or generically as “β-secretase” or “beta-secretase”), includinghomologs, isoforms and subdomains thereof that retain proteolyticactivity. Sequence identities of active memapsin 2 proteins and proteinfragments (and nucleic acid coding sequences thereof) have beenpreviously disclosed and discussed in detail in U.S. Application No.20040121947, and International Application No. PCT/US02/34324(Publication No. WO 03/039454, International publication WO 01/00663,U.S. Pat. No. 6,545,127), which are herein incorporated by reference forall purposes in their entirety.

“Amyloid beta (Aβ or Abeta)” refers to a peptide of 36-43 amino acids.While best known as a component of amyloid plaques in association withAlzheimer's disease, as Aβ is the main component of certain depositsfound in the brains of patients with Alzheimer's disease. The differentAβ isoforms (for example, Aβ40, Aβ42, and so on) refer to cleavageproducts of transmembranous APP via the β-secretase pathway. Thecleavage by β-secretase (BACE1) liberates the Aβ N-terminus, togetherwith sAPPβ and a C-terminal fragment C99. C99 is subsequently cleaved byγ-secretase to yield Aβ.

“Diseases or conditions associated with and/or mediated by β-secretaseactivity, hydrolysis of a β-secretase cleavage site of an amyloidprecursor protein, and/or β-amyloid protein accumulation” as usedherein, includes, but is not limited to, diseases such as Glaucoma, MCI(Mild cognitive impairment) or Alzheimer's disease. In anotherembodiment, the term includes, but is not limited to, MCI (Mildcognitive impairment) or Alzheimer's disease. In another embodiment, theterm includes, but is not limited to, Alzheimer's disease. In anotherembodiment, the term includes, but is not limited to, MCI (Mildcognitive impairment). In some embodiments, the compounds of formula (I)or a salt thereof can be used as agent for preventing or treatingdiseases or conditions including, but not limited to, stroke,cerebrovascular dementia, Down syndrome, Parkinson's disease (PD), anddementia with Lewy bodies (DLB).

As used herein and in the appended claims, the singular forms “a,” “an,”and “the” include plural reference unless the context clearly indicatesotherwise.

The term, “effective amount,” and cognates of this term, as used herein,refer to an amount that results in a desired pharmacological and/orphysiological effect for a specified condition (e.g., disease, disorder,etc.) or one or more of its symptoms and/or to completely or partiallyprevent the occurrence of the condition or symptom thereof and/or may betherapeutic in terms of a partial or complete cure for the conditionand/or adverse effect attributable to the condition. In reference toconditions mediated by memapsin 2 (β-secretase) or diseases orconditions associated with and/or mediated by β-secretase activity,hydrolysis of a β-secretase cleavage site of an amyloid precursorprotein, and/or β-amyloid protein accumulation, a pharmaceutically ortherapeutically effective amount comprises an amount sufficient to,among other things, cause antagonism or inhibition of memapsin 2(β-secretase). In reference to glaucoma, a pharmaceutically ortherapeutically effective amount comprises an amount sufficient to,among other things, decrease intraocular pressure; and/or halt, reverse,and/or diminish the loss of retinal ganglion cells (RGCs). In certainembodiments, the pharmaceutically effective amount is sufficient toprevent the condition, as in being administered to an individualprophylactically.

The “effective amount” will vary depending on the composition beingadministered, the condition being treated/prevented, the severity of thecondition being treated or prevented, the age and relative health of theindividual, the route and form of administration, the judgment of theattending medical or veterinary practitioner, and other factorsappreciated by the skilled artisan in view of the teaching providedherein.

The “subject” means the animal which needs its prevention or treatmentand the human who needs its prevention or treatment, in some embodimentsit means the human who needs its prevention or treatment.

When used with respect to methods of treatment/prevention and the use ofthe compounds and compositions thereof described herein, a subject “inneed thereof” may be an individual who has been diagnosed with orpreviously treated for the condition to be treated. With respect toprevention, the subject in need thereof may also be an individual who isat risk for a condition (e.g., a family history of the condition,life-style factors indicative of risk for the condition, etc.).

In some variations, the subject has been identified as having one ormore of the conditions described herein. In some embodiments, thesubject has been identified as susceptible to one or more of theconditions as described herein. The susceptibility of a subject may bebased on any one or more of a number of risk factors and/or diagnosticapproaches appreciated by the skilled artisan, including, but notlimited to, genetic profiling, family history, medical history (e.g.,appearance of related conditions), lifestyle or habits.

Examples of the embodiment of the substituent acceptable in the“R^(A11), R^(A12), R^(A21) and R^(A22) are combined with each other toform an aryl group, which is substituted” include, but are not limitedto, e.g., halogen.

Examples of the embodiment of the substituent acceptable in the “ahetero ring group, which is substituted” in B include, but are notlimited to, e.g., halogen.

Examples of the embodiment of the substituent acceptable in the“cycloalkyl, which is substituted” in B include, but are not limited to,e.g., halogen.

Examples of the embodiment of the substituent acceptable in the “loweralkyl, which is substituted” in X and Y include, but are not limited to,the groups shown in i) to iii) below.

i) halogen,

ii) cycloalkyl, or

iii) aryl.

Examples of the embodiment of the substituent acceptable in the“cycloalkyl, which is substituted” in X and Y include, but are notlimited to, the groups shown in i) to iii) below.

i) halogen,

ii) cycloalkyl, or

iii) aryl.

Examples of the embodiment of the substituent acceptable in the “X and Yare combined with each other to form a cycloalkyl group, which issubstituted” include, but are not limited to, the groups shown in i) toiii) below.

i) halogen,

ii) cycloalkyl, or

iii) aryl.

Examples of the embodiment of the substituent acceptable in the “ahetero ring group, which is substituted” in R¹, R², R³ and R⁴ include,but are not limited to, the groups shown in i) to vi) below.

i) halogen,

ii) lower alkyl, which is unsubstituted or substituted with one or moresubstituents selected from the group consisting of halogen and —O-(loweralkyl),

iii) lower alkynyl, which is unsubstituted or substituted with one ormore substituents selected from the group consisting of —O— (loweralkyl) and cycloalkyl,

iv) —O-(lower alkyl), wherein said lower alkyl is unsubstituted orsubstituted with halogen,

v) cycloalkyl, or

vi) —CN.

Examples of the embodiment of the substituent acceptable in the “loweralkyl, which is substituted” in R¹, R², R³ and R⁴ include, but are notlimited to, —O-(lower alkyl), or aryl, wherein said aryl isunsubstituted or substituted with lower alkyl.

Examples of the embodiment of the substituent acceptable in the “loweralkenyl, which is substituted” in R¹, R², R³ and R⁴ include, but are notlimited to, —O-(lower alkyl).

Examples of the embodiment of the substituent acceptable in the“—N(H)-(hetero ring group), wherein said hetero ring group issubstituted” in R¹, R², R³ and R⁴ include, but are not limited to,halogen or —O-(lower alkyl).

Examples of embodiments of substituents acceptable in the“—N(H)—C(O)-(hetero ring group), wherein said hetero ring group issubstituted” in R¹, R², R³ and R⁴ include, but are not limited to, thegroups shown in i) to vii) below.

i) halogen,

ii) lower alkyl, which is unsubstituted or substituted with one or moresubstituents selected from the group consisting of halogen, —O-(loweralkyl), and a hetero ring group,

iii) —CN,

iv) —O-(lower alkyl), wherein said lower alkyl is unsubstituted orsubstituted with halogen,

v) cycloalkyl,

vi) aryl, or

vii) a hetero ring group.

Examples of other embodiments of substituents acceptable in the“—N(H)—C(O)-(hetero ring group), wherein said hetero ring group issubstituted” in R¹, R², R³ and R⁴ include, but are not limited to, thegroups shown in i) to vii) below.

i) halogen,

ii) lower alkyl, which is unsubstituted or substituted with one or moresubstituents selected from the group consisting of halogen, —O-(loweralkyl), and a nitrogen-containing monocyclic hetero ring group,

iii) —CN,

iv) —O-(lower alkyl), wherein said lower alkyl is unsubstituted orsubstituted with halogen,

v) cycloalkyl,

vi) aryl, or

vii) a nitrogen-containing monocyclic hetero ring group.

Examples of other embodiments of the substituent acceptable in the“—N(H)—C(O)-(hetero ring group), wherein said hetero ring group issubstituted” in R¹, R², R³ and R⁴ include, but are not limited to, thegroups shown in i) to iv) below.

i) halogen,

ii) lower alkyl,

iii) —CN, or

iv) —O-(lower alkyl).

Examples of embodiments of substituents acceptable in the “cycloalkenyl,which is substituted” in R¹, R², R³ and R⁴ include, but are not limitedto, lower alkyl.

Examples of the embodiment of the substituent acceptable in the “aryl,which is substituted” in R¹, R², R³ and R⁴ include, but are not limitedto, the groups shown in i) to ix) below.

i) —OH,

ii) halogen,

iii) lower alkyl, which is unsubstituted or substituted with halogen,

iv) —O-(lower alkyl), wherein said lower alkyl is unsubstituted orsubstituted with halogen,

v) —S-(lower alkyl),

vi) cycloalkyl,

vii) —CN,

viii) lower alkenyl, which is unsubstituted or substituted with —CN, or

ix) —C(O)—N(H)-(lower alkyl).

Examples of the embodiment of groups of compounds of formula (I) of thepresent invention are shown below.

(1)

(1-1)

A¹ is O or S;

A² is —C(R^(A21)R^(A22))—; and

R^(A21) and R^(A22) are H.

(1-2)

(1-2-1)

A¹ is O;

A² is —C(R^(A21)R^(A22))—; and

R^(A21) and R^(A22) are H.

(2)

(2-1)

(2-1-1)

B is a hetero ring group, wherein said hetero ring group isunsubstituted or substituted with halogen, or cycloalkyl, wherein saidcycloalkyl is unsubstituted or substituted with halogen.

(2-1-2)

B is a hetero ring group or cycloalkyl, wherein said cycloalkyl isunsubstituted or substituted with halogen.

(2-1-3)

B is an oxygen-containing monocyclic saturated hetero ring group orcycloalkyl, wherein said cycloalkyl is unsubstituted or substituted withhalogen.

(2-1-4)

B is an oxygen-containing monocyclic saturated hetero ring group or C₃₋₆cycloalkyl, wherein said C₃₋₆ cycloalkyl is unsubstituted or substitutedwith halogen.

(2-1-5)

B is an oxygen-containing monocyclic saturated hetero ring group or C₃₋₆cycloalkyl, wherein said C₃₋₆ cycloalkyl is unsubstituted or substitutedwith F.

(2-1-6)

B is a cyclic ether group or cycloalkyl, wherein said cycloalkyl isunsubstituted or substituted with halogen.

(2-1-7)

B is a cyclic ether group or C₃₋₆ cycloalkyl, wherein said C₃₋₆cycloalkyl is unsubstituted or substituted with halogen.

(2-1-8)

B is a cyclic ether group or C₃₋₆ cycloalkyl, wherein said C₃₋₆cycloalkyl is unsubstituted or substituted with F.

(2-1-9)

B is oxetanyl, tetrahydropyranyl, cyclopropyl, cyclobutyl, or3,3-difluorocyclobutan-1-yl.

(2-1-10)

B is oxetanyl, tetrahydrofuranyl, cyclopropyl, cyclobutyl, or3,3-difluorocyclobutan-1-yl.

(2-2)

(2-2-1)

B is a hetero ring group.

(2-2-2)

B is an oxygen-containing monocyclic saturated hetero ring group.

(2-2-3)

B is a cyclic ether group.

(2-2-4)

B is oxetanyl or tetrahydropyranyl.

(2-2-5)

B is oxetanyl.

(2-2-6)

B is oxetanyl or tetrahydrofuranyl.

(2-3)

(2-3-1)

B is cycloalkyl, wherein said cycloalkyl is unsubstituted or substitutedwith halogen.

(2-3-2)

B is C₃₋₆ cycloalkyl, wherein said C₃₋₆ cycloalkyl is unsubstituted orsubstituted with halogen.

(2-3-3)

B is C₃₋₆ cycloalkyl, wherein said C₃₋₆ cycloalkyl is unsubstituted orsubstituted with F.

(2-3-4)

B is cyclopropyl, cyclobutyl, or 3,3-difluorocyclobutan-1-yl.

(2-3-5)

B is cyclopropyl.

(3)

(3-1)

X is lower alkyl; and

Y is lower alkyl.

(3-2)

X is methyl; and

Y is methyl.

(4)

(4-1)

(4-1-1)

R¹, R², R³ and R⁴ are independently selected from the group consistingof

H,

halogen, and

—N(H)—C(O)-(hetero ring group), wherein said hetero ring group isunsubstituted or substituted with one or more substituents selected fromthe group consisting of halogen,

lower alkyl which is unsubstituted or substituted with halogen,

—CN, and

—O-(lower alkyl).

(4-1-2)

R¹ and R⁴ are H;

R³ is H or halogen; and

R² is —N(H)—C(O)-(hetero ring group), wherein said hetero ring group isunsubstituted or substituted with one or more substituents selected fromthe group consisting of

halogen,

lower alkyl which is unsubstituted or substituted with halogen,

—CN, and

—O-(lower alkyl).

(4-1-3)

R¹ and R⁴ are H;

R³ is H or halogen; and

R² is —N(H)—C(O)-(nitrogen-containing monocyclic hetero ring group),wherein said nitrogen-containing monocyclic hetero ring group isunsubstituted or substituted with one or more substituents selected fromthe group consisting of halogen,

lower alkyl which is unsubstituted or substituted with halogen,

—CN, and

—O-(lower alkyl).

(4-1-4)

R¹ and R⁴ are H;

R³ is H or halogen; and

R² is selected from the group consisting of

—N(H)—C(O)-(pyridyl), wherein said pyridyl is unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of

halogen,

lower alkyl which is unsubstituted or substituted with halogen,

—CN, and

—O-(lower alkyl),

—N(H)—C(O)-(pyrazinyl), wherein said pyrazinyl is unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of

halogen,

lower alkyl which is unsubstituted or substituted with halogen,

—CN, and

—O-(lower alkyl), and

—N(H)—C(O)-(pyrimidinyl), wherein said pyrimidinyl is unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of

halogen,

lower alkyl which is unsubstituted or substituted with halogen,

—CN, and

—O-(lower alkyl).

(4-1-5)

R¹ and R⁴ are H;

R³ is H or halogen; and

R² is selected from the group consisting of

—N(H)—C(O)-(pyridyl), wherein said pyridyl is unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of

halogen,

lower alkyl, and

—CN,

—N(H)—C(O)-(pyrazinyl), wherein said pyrazinyl is unsubstituted orsubstituted with —O-(lower alkyl) or

lower alkyl which is unsubstituted or substituted with halogen, and

—N(H)—C(O)-(pyrimidinyl), wherein said pyrimidinyl is unsubstituted orsubstituted with halogen.

(4-1-6)

R¹ and R⁴ are H;

R³ is H or halogen; and

R² is —N(H)—C(O)-(pyridyl), wherein said pyridyl is unsubstituted orsubstituted with one or more substituents selected from the groupconsisting of

halogen,

lower alkyl, and

—CN.

(4-1-6-1)

R¹ and R⁴ are H;

R³ is H or halogen; and

R² is —N(H)—C(O)-(pyridyl), wherein said pyridyl is unsubstituted orsubstituted with halogen.

(4-1-7)

R¹ and R⁴ are H;

R³ is H or halogen; and

R² is —N(H)—C(O)-(pyrazinyl), wherein said pyrazinyl is unsubstituted orsubstituted with —O-(lower alkyl) or

lower alkyl which is unsubstituted or substituted with halogen.

(4-1-8)

R¹ and R⁴ are H;

R³ is H or halogen; and

R² is —N(H)—C(O)-(pyrimidinyl), wherein said pyrimidinyl isunsubstituted or substituted with halogen.

(4-2)

The groups of any one of (4-1),

wherein R³ is H.

Furthermore, still other embodiments of the compounds of formula (I) ofthe present invention include the compounds including a combination oftwo or more of the groups described in (1) to (4) above, specifically,the following compounds.

(5) The compound of formula (I), wherein B is as described in (2).

(6) The compound as described in (5), wherein X and Y are as describedin (3).

(7) The compound as described in (5) or (6), wherein R¹, R², R³ and R⁴are as described in (4).

(8) The compound as described in (5), (6) or (7), wherein A¹ and A² areas described in (1).

Furthermore, still other embodiments of the compounds of formula (I) ofthe present invention include the compounds including a combination oftwo or more of the groups described in (1) to (4) above, specifically,the following compounds.

(9) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-1-3), X and Y are as described in (3-1)and R¹, R², R³ and R⁴ are as described in (4-1-2).

(10) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-1-3), X and Y are as described in (3-1)and R¹, R², R³ and R⁴ are as described in (4-1-5).

(11) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-2), X and Y are as described in (3-1)and R¹, R², R³ and R⁴ are as described in (4-1-5).

(12) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-1), X and Y are as described in (3-1)and R¹, R², R³ and R⁴ are as described in (4-1-5).

(13) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-1)and R¹, R², R³ and R⁴ are as described in (4-1-5).

(14) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-5).

(15) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-6).

(16) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-6-1).

(17) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-7).

(18) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-8).

(19) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-6), wherein R³ is H.

(20) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-6-1), wherein R³ is H.

(21) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-7), wherein R³ is H.

(22) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-8), wherein R³ is H.

(23) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-1)and R¹, R², R³ and R⁴ are as described in (4-1-5).

(24) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-5).

(25) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-6).

(26) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-6-1).

(27) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-7).

(28) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-8).

(29) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-6), wherein R³ is H.

(30) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-6-1), wherein R³ is H.

(31) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-7), wherein R³ is H.

(32) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2)and R¹, R², R³ and R⁴ are as described in (4-1-8), wherein R³ is H.

(33) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-1-3), X and Y are as described in (3-1)and R¹, R², R³ and R⁴ are as described in (4-1-3).

(34) The compound of formula (I), wherein A¹ and A² are as described in(1-2-1), B is as described in (2-1-3), X and Y are as described in (3-1)and R¹, R², R³ and R⁴ are as described in (4-1-4).

(35) The compound as described in (5) to (34), wherein the compound offormula (I) is the compound of formula (II) as below.

Examples of the specific compounds encompassed by the present inventioninclude the following compounds. Nomenclature of some compoundsdescribed herein may be identified using IUPAC or other namingconventions including ACD/Name ver. 12.02, available from AdvancedChemistry Development, Inc., Toronto, Ontario, Canada.

-   N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-chloropyridine-2-carboxamide,-   N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-fluoropyridine-2-carboxamide,-   N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-chloro-3-fluoropyridine-2-carboxamide,-   N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-bromopyrimidine-2-carboxamide,-   N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-chloro-3-methylpyridine-2-carboxamide,-   N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-cyanopyridine-2-carboxamide,-   N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-methoxypyrazine-2-carboxamide,-   N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide,-   N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide,-   N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide,-   N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide,-   N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-bromopyridine-2-carboxamide,-   N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-chloropyridine-2-carboxamide,    and-   N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-fluoropyridine-2-carboxamide.

Other examples of the specific compounds encompassed by the presentinvention include the following compounds. Nomenclature of somecompounds described herein may be identified using IUPAC or other namingconventions including ACD/Name ver. 12.02, available from AdvancedChemistry Development, Inc., Toronto, Ontario, Canada.

-   N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide,-   N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide,    and-   N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide.

The present invention relates to a hydrate of the compound or a salt,wherein said compound is

-   N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide.

In some embodiments,

A¹ is O;

A² is CH₂;

B is

or cyclopropyl;

X and Y are both methyl, or X and Y are both H;

R¹, R³, and R⁴ are H; and

R² is —N(H)—C(O)-(hetero ring group), wherein the hetero ring group ofR² is selected from the group consisting of

each of which is substituted with 1 or 2 substituents independentlyselected from the group consisting of halogen, cyano, unsubstituted—O-lower alkyl, unsubstituted lower alkyl, lower alkyl substituted withone or more halogen, —O-lower alkyl substituted with one or morehalogen, lower alkyl substituted with —OCH₃, unsubstituted loweralkynyl, and unsubstituted cycloalkyl. In some embodiments, B is

and X and Y are both methyl. In some embodiments, B is cyclopropyl, andX and Y are both H. In some embodiments, R² is selected from the groupconsisting of

and wherein R⁵ and R⁶ are independently selected from the groupconsisting of hydrogen, fluoro, chloro, bromo, cyano, —OCH₃, methyl,—CHF₂, —OCHF₂, —OCH₂CHF₂, —CH₂OCH₃, —C≡C—CH₃, and cyclopropyl. In someembodiments, R⁵ is chloro or —OCH₃, and R⁶ is hydrogen or fluoro. Insome embodiments, R⁵ is —CHF₂ or —OCHF₂, and R⁶ is hydrogen.

In some embodiments,

A¹ is O;

A² is CH₂; and

R² is —N(H)—C(O)-(hetero ring group), wherein the hetero ring group is

In some embodiments, the compound is selected from the group consistingof

or a salt thereof.

In some embodiments, the compound is

or a salt thereof.

In some embodiments, the compound is

or a salt thereof.

In some embodiments, the compound is

or a salt thereof.

In some embodiments, the compound is selected from the group consistingof

or a salt thereof.

In some embodiments, the compound is

or a salt thereof.

In some embodiments, the compound is

or a salt thereof.

In some embodiments, the compound is

or a salt thereof.

In some embodiments, the compound is selected from the group consistingof

or a salt thereof.

The compound of the formula (I) may exist in the form of tautomers orgeometrical isomers depending on the kind of substituents. In thepresent specification, the compound of the formula (I) shall bedescribed in only one form of isomer, yet the present invention includessuch an isomer, isolated forms of the isomers, or a mixture thereof.

In addition, the compound of the formula (I) may have asymmetric carbonatoms or axial asymmetry in some cases, and correspondingly, it mayexist in the form of optical isomers. The present invention includesboth an isolated form of the optical isomers of the compound of theformula (I) or a mixture thereof.

Moreover, the present invention also includes a pharmaceuticallyacceptable prodrug of the compound of the formula (I). Thepharmaceutically acceptable prodrug is a compound having a group thatcan be converted into an amino group, a hydroxyl group, a carboxylgroup, or the like through solvolysis or under physiological conditions.Examples of the group forming the prodrug include the groups describedin Prog. Med., 5, 2157-2161 (1985) and Pharmaceutical Research andDevelopment, Drug Design, Hirokawa Publishing Company (1990), Vol. 7,163-189, which is incorporated by reference herein in its entirety.

Furthermore, the salt of the compound of the formula (I) is apharmaceutically acceptable salt of the compound of the formula (I) andmay form an acid addition salt or a salt with a base depending on thekind of substituents. Specific examples thereof include acid additionsalts with inorganic acids such as hydrochloric acid, hydrobromic acid,hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and thelike, and with organic acids such as formic acid, acetic acid, propionicacid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleicacid, lactic acid, malic acid, mandelic acid, tartaric acid,dibenzoyltartaric acid, ditoluoyltartaric acid, citric acid,methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid,p-toluenesulfonic acid, aspartic acid, glutamic acid, and the like, andsalts with inorganic bases such as sodium, potassium, magnesium,calcium, aluminum, and the like or organic bases such as methylamine,ethylamine, ethanolamine, lysine, ornithine, and the like, salts withvarious amino acids or amino acid derivatives such as acetylleucine andthe like, ammonium salts, etc.

In addition, the present invention also includes various hydrates orsolvates, and polymorphic crystal substances of the compound of theformula (I) and a salt thereof. In addition, the present invention alsoincludes compounds labeled with various radioactive or non-radioactiveisotopes.

(Preparation Methods)

The compound of the formula (I) and a salt thereof can be prepared usingthe characteristics based on the basic structure or the type ofsubstituents thereof and by applying various known synthesis methods.During the preparation, replacing the relevant functional group with asuitable protective group (a group that can be easily converted into thefunctional group) at the stage from starting material to an intermediatemay be effective depending on the type of the functional group in theproduction technology in some cases. The protective group for such afunctional group may include for example, the protective groupsdescribed in “Greene's Protective Groups in Organic Synthesis (4^(th)Ed., 2006)”, P. G. M. Wuts and T. W. Greene, and one of these may beselected and used as necessary depending on the reaction conditions. Inthis kind of method, a desired compound can be obtained by introducingthe protective group, by carrying out the reaction and by eliminatingthe protective group as necessary.

In addition, the prodrug of the compound of the formula (I) can beproduced by introducing a specific group or by carrying out the reactionusing the obtained compound of the formula (I) at the stage from astarting material to an intermediate, just as in the case of theabove-mentioned protective group. The reaction can be carried out usingmethods known to those skilled in the art, such as ordinaryesterification, amidation, dehydration, and the like.

Hereinbelow, the representative preparation methods for the compound ofthe formula (I) will be described. Each of the production processes mayalso be carried out with reference to the References appended in thepresent description. Unless otherwise indicated, in any of the foregoingschemes, A¹, A², R¹, R², R³, R⁴, X, Y, and B are as described forformula (I) or any applicable variation thereof. Further, thepreparation methods of the compound of the formula (I) are not limitedto the examples as shown below.

(Production Process 1)

A compound (I) can be obtained by subjecting a compound (15) to additionand cyclization reactions.

In the addition reaction, the compound (15) and an equivalent amount oran excess amount of iodine and silver cyanate or silver thiocyanate areused, and a mixture thereof is stirred under any temperature conditionfrom cooling to heating and refluxing, preferably at 0° C. to 200° C.,and still more preferably at 20° C. to 120° C., usually for 0.1 hours to5 days, in a solvent which is inert to the reaction or without asolvent. Examples of the solvent as used herein are not particularlylimited, but include alcohols such as methanol, ethanol, tert-butanol,and the like, aromatic hydrocarbons such as benzene, toluene, xylene,and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane,dimethoxyethane, and the like, halogenated hydrocarbons such asdichloromethane, 1,2-dichloroethane, chloroform, and the like,N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile,and a mixture thereof.

In the cyclization reaction, the crude mixture after the additionreaction and an equivalent amount or an excess amount of NH₃ dissolvedin solvent such as H₂O or ethanol (EtOH), and so on, are used, and amixture thereof is stirred under any temperature condition from coolingto heating to refluxing, preferably at 0° C. to 200° C., and still morepreferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in asolvent which is inert to the reaction or without a solvent. Examples ofthe solvent as used herein are not particularly limited, but includealcohols such as methanol, ethanol, tert-butanol, and the like, aromatichydrocarbons such as benzene, toluene, xylene, and the like, ethers suchas diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and thelike, halogenated hydrocarbons such as dichloromethane,1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide,dimethylsulfoxide, ethyl acetate, acetonitrile, water and a mixturethereof. It may be advantageous in some cases for the smooth progress ofthe reaction to carry out the reaction in the presence of an organicbase such as triethylamine, N,N-diisopropylethylamine,N-methylmorpholine, and the like, or an inorganic base such as sodiumtert-butoxide, potassium carbonate, sodium bis(trimethylsilyl)amide,sodium carbonate, potassium hydroxide, and the like.

In some embodiments of Production Process 1, the compound (I) is acompound of formula (Ie):

wherein A¹¹ represents O or S, and A²¹ represents CH₂.(Production Process 2)

A compound (I) can be obtained by subjecting a compound (16) and NH₃ toa substitution reaction.

In this reaction, the compound (16) and an equivalent amount or anexcess amount of NH₃ dissolved in solvent such as H₂O or EtOH, and soon, are used, and a mixture thereof is stirred under any temperaturecondition from cooling to heating and refluxing, preferably at 0° C. to200° C., and still more preferably at 20° C. to 120° C., usually for 0.1hours to 5 days, in a solvent which is inert to the reaction or withouta solvent. It may be advantageous in some cases for the smooth progressof the reaction to carry out the reaction under microwave irradiation.Examples of the solvent as used herein are not particularly limited, butinclude alcohols such as methanol, ethanol, tert-butanol, and the like,aromatic hydrocarbons such as benzene, toluene, xylene, and the like,ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane,and the like, halogenated hydrocarbons such as dichloromethane,1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide,dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof.It may be advantageous in some cases for the smooth progress of thereaction to carry out the reaction in the presence of tert-butylhydroperoxide, and the like.

(Other Production Processes)

Furthermore, several substituents in the formula (I) can also be easilyconverted into other functional groups by using the compound of thepresent invention (I) as a starting material by means of the reactionsapparent to a person skilled in the art, or modified methods thereof.The reaction can be carried out by any combination of the processes thatcan be usually employed by a person skilled in the art, such ashydrolysis, alkylation, halogenation, hydrogenation, and the like.Several examples thereof are presented below.

(Production Process 3)

(wherein R^(B) represents H or lower alkyl, or two R^(B)'s are combinedwith each other to form C₂₋₇ alkylene, Lv represents a leaving group,and R^(2a) represents a group in R² with the exception that R^(2a)cannot be H or halogen. In some embodiments, R^(2a) represents aryl or ahetero ring group which has aromaticity in R². Moreover, said aryl maybe substituted with substituents acceptable in the “aryl” of R¹, R², R³and R⁴, and said hetero ring group may be substituted with substituentsacceptable in the “a hetero ring group” of R¹, R², R³ and R⁴.).

First, the compound (17) can be obtained by subjecting the compound (Ia)to a cross-coupling reaction with a borylation reagent.

In this reaction, a mixture of the compound (Ia) and a borylationreagent in equivalent amounts, or with either thereof in an excessamount is stirred under any temperature condition from cooling toheating, and preferably −20° C. to 60° C., usually for 0.1 hours to 5days, in a solvent which is inert to the reaction, in the presence of anorganometallic compound. The solvent as used herein is not particularlylimited, but examples thereof include aromatic hydrocarbons such asbenzene, toluene or xylene, and the like, halogenated hydrocarbons suchas dichloromethane, 1,2-dichloroethane, chloroform, and the like, etherssuch as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, andthe like, DMF, DMSO, EtOAc, acetonitrile, water, and a mixture thereof.Examples of the borylation reagent include bis(pinacolato)diboron, andthe like. It may be advantageous in some cases for the smooth progressof the reaction to carry out the reaction in the presence of inorganicbase, such as potassium acetate or potassium phenolate, and the like.Examples of an organometallic compound include palladium catalysts, suchas [1,1′-Bis(diphenylphosphino)ferrocene]palladium(II) dichloride. Itmay be advantageous in some cases for the smooth progress of thereaction to carry out the reaction after protecting —NH₂ of a compound(Ia).

Moreover, the compound (Ib) can be obtained by subjecting the compound(17) and the R^(2a)-Lv to a coupling reaction. Herein, examples of theleaving group Lv include halogen, a trifluoromethanesulfonyloxy group,and the like.

In this reaction, a mixture of the compound (17) and an equivalentamount or an excess amount of R^(2a)-Lv is stirred under any temperaturecondition from cooling to heating and refluxing, and preferably at 0° C.to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inertto the reaction or without a solvent by using a catalyst used forSuzuki-Miyaura cross-coupling reaction. The solvent as used herein isnot particularly limited, but examples thereof include aromatichydrocarbons such as benzene, toluene, xylene, and the like, ethers suchas dimethyl ether, diethyl ether, tetrahydrofuran, dioxane,dimethoxyethane, and the like, halogenated hydrocarbons such asdichloromethane, 1,2-dichloroethane, chloroform, and the like,N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile,and a mixture thereof. The catalyst as used herein is not particularlylimited, but tetrakis(triphenylphosphine)palladium(0), palladium(II)acetate, dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II),bis(triphenylphosphine)palladium(II) chloride,tris(dibenzylideneacetone)dipalladium(0)-2-dicyclohexylphosphino-2′,6′-dimethoxybiphenylor the like can be used. In addition, metal palladium(0) can also beused to carry out the coupling reaction. It may be advantageous in somecases for the smooth progress of the reaction to carry out the reactionafter protecting —NH₂ of a compound (17).

(Production Process 4)

A compound (Ib) can be obtained by subjecting a compound (Ia) andR²—B(OR^(B))₂ to a coupling reaction. This reaction can be conducted bythe same condition of the said reaction of (Production Process 3). Itmay be advantageous in some cases for the smooth progress of thereaction to carry out the reaction after protecting —NH₂ of a compound(Ia).

(Production Process 5)

(wherein R²² represents a hetero ring group which may be substitutedwith substituents acceptable in the “—N(H)-(hetero ring group)” of R¹,R², R³ and R⁴.).

A compound (Ic) among the compounds (I) of the present invention can beobtained by subjecting a compound (Ia) and NR²²H₂ to a substitutionreaction.

In this reaction, the compound (Ia) and an equivalent amount or anexcess amount of NR²²H₂ are used, and a mixture thereof is stirred underany temperature condition from cooling to heating and refluxing,preferably at 0° C. to 200° C., and still more preferably at 20° C. to120° C., usually for 0.1 hours to 5 days, in a solvent which is inert tothe reaction or without a solvent. It may be advantageous in some casesfor the smooth progress of the reaction to carry out the reaction undermicrowave irradiation. Examples of the solvent as used herein are notparticularly limited, but include alcohols such as methanol, ethanol,tert-butanol, and the like, aromatic hydrocarbons such as benzene,toluene, xylene, and the like, ethers such as diethyl ether,tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenatedhydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform,and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate,acetonitrile, and a mixture thereof. It may be advantageous in somecases for the smooth progress of the reaction to carry out the reactionin the presence of an organic base such as triethylamine,N,N-diisopropylethylamine, N-methylmorpholine, and the like, or aninorganic base such as sodium tert-butoxide, potassium carbonate,caesium carbonate, sodium bis(trimethylsilyl)amide, sodium carbonate,potassium hydroxide, and the like. It may be advantageous in some casesfor the smooth progress of the reaction to carry out the reaction afterprotecting —NH₂ of a compound (Ia).

Moreover, the reaction may be carried out using a catalyst which is notparticularly limited, but includes catalysts used for Ullmann reaction,Buchwald-Hartwig reaction, or the like. The catalyst forBuchwald-Hartwig reaction as used herein is not particularly limited,but a suitable combination of tris(dibenzylideneacetone)palladium (0),tetrakis(triphenylphosphine)palladium (0), or the like with4,5-bis(diphenylphosphino)-9,9′-dimethylxanthene (Xantphos),2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (SPhos),2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (XPhos), and thelike can be used. The catalyst for Ullmann reaction as used herein isnot particularly limited, but a suitable combination of copper(I)iodide, or the like with (1R*,2R*)—N,N′-dimethylcyclohexane-1,2-diamine,1,10-phenanthroline and the like can be used.

(Production Process 6)

(wherein R²¹ represents a hetero ring group which may be substitutedwith substituents acceptable in the “—N(H)—C(O)-(hetero ring group)” ofR¹, R², R³ and R⁴, Boc represents a tert-butoxylcarbonyl group.)

A compound (19) can be obtained by subjecting a compound (18) which isobtained by the protection reaction of (Ia) and lithiumbis(trimethylsilyl)amide to an amination reaction.

In this reaction, the compound (18) and an equivalent amount or anexcess amount of lithium bis(trimethylsilyl)amide are used, and amixture thereof is stirred under any temperature condition from coolingto heating and refluxing, preferably at 0° C. to 200° C., and still morepreferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in asolvent which is inert to the reaction or without a solvent in thepresence of a palladium catalyst. The palladium catalyst can be preparedin situ from bis(dibenzylideneacetone)palladium (0) andtri-tert-butylphosphonium tetrafluoroborate. Examples of the solvent asused herein are not particularly limited, but include aromatichydrocarbons such as benzene, toluene, xylene, and the like, ethers suchas diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and thelike, halogenated hydrocarbons such as dichloromethane,1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide,dimethylsulfoxide, and a mixture thereof.

A compound (20) can be obtained by subjecting a compound (19) to anamination reaction. For the reaction, the compound (19) and anequivalent amount or an excess amount of R²¹—C(═O)—OH are used, and amixture thereof is stirred in a range of from cooling to heating,preferably at a temperature from −20° C. to 60° C., usually for about0.1 hours to 5 days, in a solvent which is inert to the reaction, in thepresence of a condensing agent. The solvent as used herein is notparticularly limited, but examples thereof include aromatic hydrocarbonssuch as benzene, toluene, xylene, and the like, halogenated hydrocarbonssuch as dichloromethane, 1,2-dichloroethane, chloroform, and the like,ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane,and the like, DMF, DMSO, EtOAc, acetonitrile, or water, and a mixturethereof. Examples of the condensing agent include, but are not limitedto, CDI, diphenylphosphoryl azide, phosphorus oxychloride, WSC(Water-Soluble Carbodiimide, trademark,1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and the like), and DCC(dicyclohexylcarbodiimide). It may be in some cases preferable for thereaction to use an additive for example, 1-hydroxybenzotriazole. It isin some cases advantageous for smooth progress of the reaction to carryout the reaction in the presence of organic bases such as triethylamine,N,N-diisopropylethylamine, N-methylmorpholine, DBU, DMAP, and the like,or inorganic bases such as potassium carbonate, sodium carbonate,potassium hydroxide, and the like.

Furthermore, it is also possible to use a method in which a reactivederivative of R²¹—C(═O)—OH is used, and reacted with the compound (19).Examples of the reactive derivative of the carboxylic acid include acidhalides that can be obtained by the reaction with a halogenating agentsuch as phosphorus oxychloride, thionyl chloride, and the like, mixedacid anhydrides that can be obtained by the reaction with isobutylchloroformate or the like, activated esters that can be obtained bycondensation with 1-hydroxybenzotriazole or the like, etc. The reactionof the reactive derivative with the compound (19) can be carried out ina range of from cooling to heating, and preferably from −20° C. to 60°C., in a solvent which is inert to the reaction, such as halogenatedhydrocarbons, aromatic hydrocarbons, ethers, and the like.

A compound (Id) can be obtained by subjecting a compound (20) to adeprotection reaction. The deprotection reaction can be carried out withreference to, for example, “Greene's Protective Groups in OrganicSynthesis (4^(th) Ed., 2006)”, P. G. M. Wuts and T. W. Greene.

Moreover, each reaction in (Production Process 6) can be conducted withusing a compound protected by protective group except a Boc group, andeach reaction may be conducted with using a non-protective compound.

(Starting Material Synthesis 1)

A compound (2) can be obtained by subjecting a compound (I) andX—(C═O)—Y to a cyclization reaction.

In this reaction, the compound (I) and an equivalent amount or an excessamount of X—(C═O)—Y are used, and a mixture thereof is stirred under anytemperature condition from cooling to heating and refluxing, preferablyat 0° C. to 200° C., and still more preferably at 20° C. to 120° C.,usually for 0.1 hours to 5 days, in a solvent which is inert to thereaction or without a solvent. Examples of the solvent as used hereinare not particularly limited, but include alcohols such as methanol,ethanol, tert-butanol, and the like, aromatic hydrocarbons such asbenzene, toluene, xylene, and the like, ethers such as diethyl ether,tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenatedhydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform,and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate,acetonitrile, and a mixture thereof. It may be advantageous in somecases for the smooth progress of the reaction to carry out the reactionin the presence of an organic acid or organic base. Examples of theorganic acid as used herein are not particularly limited, but includeacetic acid, trifluoroacetic acid and the like, and examples of theorganic base as used herein are not particularly limited, but includepyrrolidine, triethylamine, N,N-diisopropylethylamine,N-methylmorpholine, and the like, or an inorganic base such as sodiumtert-butoxide, potassium carbonate, sodium bis(trimethylsilyl)amide,sodium carbonate, potassium hydroxide, caesium carbonate, and the like.

A compound (3) can be obtained by subjecting a compound (2) and HCHO orLv-CH₂—OH to a substitution reaction in the presence of a base. Herein,examples of the leaving group Lv include a benzotriazolyl group, and thelike.

In this reaction, the compound (2) and an equivalent amount or an excessamount of HCHO or Lv-CH₂—OH are used, and a mixture thereof is stirredunder any temperature condition from cooling to heating and refluxing,preferably at 0° C. to 200° C., and still more preferably at 20° C. to120° C., usually for 0.1 hours to 5 days, in a solvent which is inert tothe reaction or without a solvent. Examples of the solvent as usedherein are not particularly limited, but include alcohols such asmethanol, ethanol, tert-butanol, and the like, aromatic hydrocarbonssuch as benzene, toluene, xylene, and the like, ethers such as diethylether, tetrahydrofuran, dioxane, dimethoxyethane, and the like,halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane,chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide,ethyl acetate, acetonitrile, water, and a mixture thereof. Examples ofthe organic base as used herein are not particularly limited, butinclude pyrrolidine, triethylamine, N,N-diisopropylethylamine,N-methylmorpholine, and the like, or an inorganic base such as sodiumtert-butoxide, potassium carbonate, sodium bis(trimethylsilyl)amide,calcium hydroxide, sodium carbonate, potassium hydroxide, caesiumcarbonate, and the like.

A compound (4) can be obtained by subjecting a compound (3) to amodified Mitsunobu reaction as described in Warren et al. J. Chem. Soc.,Perkin Trans. 1, 2001, 2983.

In this reaction, a compound (3) is treated under any temperaturecondition from cooling to heating, and preferably −20° C. to 80° C.,usually for 0.1 hours to 3 days, in a solvent which is inert to thereaction, in the presence of zinc bis(dimethyldithiocarbamate), an azocompound and a phosphorous compound. Examples of the solvent as usedherein are not particularly limited, but include ethers such as diethylether, tetrahydrofuran, dioxane, dimethoxyethane, and the like,halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane,chloroform, and the like, aromatic hydrocarbons such as benzene,toluene, xylene, and the like, N,N-dimethylformamide, dimethylsulfoxide,and a mixture thereof. As the azo compound, diesters of azodicarboxylicacid, such as, diethyl azodicarboxylate, or diisopropyl azodicarboxylatecan be used, and as the phosphorous compound, for example,triphenylphosphine is suitably used.

(Starting Material Synthesis 2)

A compound (6) can be obtained by subjecting the compound (5) to Wittigreaction.

In this reaction, the compound (5) is treated under any temperaturecondition from cooling to heating, and preferably −20° C. to 80° C.,usually for 0.1 hours to 3 days, in a solvent which is inert to thereaction, in the presence of an equivalent amount or an excess amount ofmethyltriphenylphosphonium halide such as methyltriphenylphosphoniumbromide in the presence of a base. Examples of the solvent as usedherein are not particularly limited, but include ethers such as diethylether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, aromatichydrocarbons such as benzene, toluene, xylene, and the like,N,N-dimethylformamide, dimethylsulfoxide, and a mixture thereof.Examples of the base as used herein are not particularly limited, butinclude sodium bis(trimethylsilyl)amide, n-butyllithium, potassiumtert-butoxide, sodium ethoxide, sodium methoxide, sodium hydride, andthe like.

(Starting Material Synthesis 3)

(wherein R^(Proc) represents a protective group, B¹ represents acycloalkyl group which may be substituted).

A compound (9) can be obtained by subjecting a compound (7) and acompound (8) to a substitution reaction. Herein, the reaction isconducted after converting —OH group of a compound (7) to a leavinggroup, such as mesyloxy group (methanesulfonyloxy group).

In this reaction, a mesylate derivative of the compound (7) and anequivalent amount or an excess amount of compound (8) are used, and amixture thereof is stirred under any temperature condition from coolingto heating and refluxing, preferably at 0° C. to 200° C., and still morepreferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in asolvent which is inert to the reaction or without a solvent. Examples ofthe solvent as used herein are not particularly limited, but includearomatic hydrocarbons such as benzene, toluene, xylene, and the like,ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane,and the like, halogenated hydrocarbons such as dichloromethane,1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide,dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof.It may be advantageous in some cases for the smooth progress of thereaction to carry out the reaction in the presence of an acid or a base.Examples of the organic acid as used herein are not particularlylimited, but include, acetic acid, trifluoroacetic acid,p-toluenesulfonic acid, methanesulfonic acid and the like, examples ofthe inorganic acid as used herein are not particularly limited, butinclude, hydrochloric acid, sulfuric acid, potassium hydrogen sulfateand the like, and examples of the organic base as used herein are notparticularly limited, but include, pyridine, 2,6-lutidine(2,6-dimethylpyridine), triethylamine, diisopropylethylamine,1,8-diazabicyclo[5.4.0]undec-7-ene and the like, examples of theinorganic base as used herein are not particularly limited, but include,sodium carbonate, potassium carbonate, sodium bicarbonate, potassiumphosphate, caesium carbonate and the like.

A compound (10) can be obtained by converting an ester group of acompound (9) to a carboxylic acid group by a hydrolysis reaction, andthen performing a cyclization reaction.

First, the hydrolysis reaction can be carried out with reference to, forexample, “Greene's Protective Groups in Organic Synthesis (4^(th) Ed.,2006)”, P. G. M. Wuts and T. W. Greene.

Next, the hydrolysis product obtained from the compound (9) is stirredunder any temperature condition from cooling to heating and refluxing,and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in asolvent which is inert to the reaction or without a solvent. Examples ofthe solvent as used herein are not particularly limited, but includearomatic hydrocarbons such as benzene, toluene, xylene, and the like,ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane,and the like, halogenated hydrocarbons such as dichloromethane,1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide,dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof.It is in some cases advantageous in advancing the reaction smoothly tocarry out the reaction under an acidic condition. Examples of the acidas used herein are not particularly limited, but include organic acidssuch as p-toluenesulfonic acid, acetic acid, and the like, and inorganicacids such as hydrochloric acid, sulfuric acid, and the like.

(Starting Material Synthesis 4)

A compound (12) can be obtained by hydrolysis reaction.

In this reaction, the compound (11) is stirred under any temperaturecondition from cooling to heating and refluxing, and preferably at 0° C.to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inertto the reaction or without a solvent in the presence of a base such aslithium hydroxide. Examples of the solvent as used herein are notparticularly limited, but include aromatic hydrocarbons such as benzene,toluene, xylene, and the like, ethers such as diethyl ether,tetrahydrofuran, dioxane, dimethoxyethane, and the like, alcohols suchas methanol, ethanol, tert-butanol, and the like, N,N-dimethylformamide,dimethylsulfoxide, acetonitrile, water and a mixture thereof.

A compound (13) can be obtained by subjecting a compound (12) andLv-C(═O)—CH₂-Lv to a substitution reaction and cyclization reaction.

In the substitution reaction, the compound (12) and an equivalent amountor an excess amount of Lv-C(═O)—CH₂-Lv are used, and a mixture thereofis stirred under any temperature condition from cooling to heating andrefluxing, preferably at 0° C. to 200° C., and still more preferably at20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent whichis inert to the reaction or without a solvent. Examples of the solventas used herein are not particularly limited, but include aromatichydrocarbons such as benzene, toluene, xylene, and the like, ethers suchas diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and thelike, halogenated hydrocarbons such as dichloromethane,1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide,dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof.It may be advantageous in some cases for the smooth progress of thereaction to carry out the reaction in the presence of an acid or base.Examples of the organic acid as used herein are not particularlylimited, but include, acetic acid, trifluoroacetic acid,p-toluenesulfonic acid, methanesulfonic acid and the like, examples ofthe inorganic acid as used herein are not particularly limited, butinclude, hydrochloric acid, sulfuric acid, potassium hydrogen sulfateand the like, and examples of the organic base as used herein are notparticularly limited, but include, pyridine, 2,6-lutidine(2,6-dimethylpyridine), triethylamine, diisopropylethylamine,1,8-diazabicyclo[5.4.0]undec-7-ene and the like, examples of theinorganic base as used herein are not particularly limited, but include,sodium carbonate, potassium carbonate, sodium bicarbonate, potassiumphosphate, caesium carbonate and the like.

In the cyclization reaction, the compound after substitution reaction ofa compound (12) is stirred under any temperature condition from coolingto heating and refluxing, preferably at 0° C. to 200° C., and still morepreferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in asolvent which is inert to the reaction or without a solvent. Examples ofthe solvent as used herein are not particularly limited, but includealcohols such as methanol, ethanol, tert-butanol, 2-methylbutan-2-ol,and the like, aromatic hydrocarbons such as benzene, toluene, xylene,and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane,dimethoxyethane, and the like, halogenated hydrocarbons such asdichloromethane, 1,2-dichloroethane, chloroform, and the like,N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile,and a mixture thereof. It may be advantageous in some cases for thesmooth progress of the reaction to carry out the reaction in thepresence of an acid or base. Examples of the organic acid as used hereinare not particularly limited, but include, acetic acid, trifluoroaceticacid, p-toluenesulfonic acid, methanesulfonic acid and the like,examples of the inorganic acid as used herein are not particularlylimited, but include, hydrochloric acid, sulfuric acid, potassiumhydrogen sulfate and the like, and examples of the organic base as usedherein are not particularly limited, but include, pyridine, 2,6-lutidine(2,6-dimethylpyridine), triethylamine, diisopropylethylamine, potassiumtert-butoxide, 1,8-diazabicyclo[5.4.0]undec-7-ene and the like, examplesof the inorganic base as used herein are not particularly limited, butinclude, sodium carbonate, potassium carbonate, sodium bicarbonate,potassium phosphate, caesium carbonate and the like.

A compound (14) can be obtained by a reaction of a compound (13) andLawesson's reagent.

In this reaction, the compound (13) is stirred under any temperaturecondition from cooling to heating and refluxing, and preferably at 0° C.to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inertto the reaction or without a solvent in the presence of Lawesson'sreagent(2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane-2,4-disulfide).Examples of the solvent as used herein are not particularly limited, butinclude aromatic hydrocarbons such as benzene, toluene, xylene, and thelike, ethers such as diethyl ether, tetrahydrofuran, dioxane,dimethoxyethane, and the like, halogenated hydrocarbons such asdichloromethane, 1,2-dichloroethane, chloroform, and the like,dimethylsulfoxide, acetonitrile, and a mixture thereof.

(Starting Material Synthesis 5)

A compound (21) can be obtained by Mannich reaction and eliminationreaction of compound (2).

In this reaction, a mixture of the compound (2),N,N,N′,N′-tetramethylmethanediamine, and acetic acid is stirred underany temperature condition from cooling to heating and refluxing, andpreferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in asolvent which is inert to the reaction or without a solvent. Examples ofthe solvent as used herein are not particularly limited, but includearomatic hydrocarbons such as benzene, toluene, xylene, and the like,ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane,and the like, halogenated hydrocarbons such as dichloromethane,1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide,dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof.Subsequently, acetic anhydride is added to the mixture and the mixtureis stirred under any temperature condition from cooling to heating andrefluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5days, in a solvent which is inert to the reaction or without a solvent.Examples of the solvent as used herein are described above.

A compound (22) can be obtained by subjecting a compound (21) to aCorey-Chaykovsky type reaction.

In this reaction, the compound (21) and an equivalent amount or anexcess amount of trimethylsulfoxonium iodide are used, and a mixturethereof is stirred under any temperature condition from cooling toheating and refluxing, preferably at 0° C. to 200° C., and still morepreferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in asolvent which is inert to the reaction or without a solvent in thepresence of a base. Examples of the solvent as used herein are notparticularly limited, but include alcohols such as methanol, ethanol,tert-butanol, and the like, aromatic hydrocarbons such as benzene,toluene, xylene, and the like, ethers such as diethyl ether,tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenatedhydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform,and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate,acetonitrile, and a mixture thereof. It may be advantageous in somecases for the smooth progress of the reaction to use a pre-formedmixture of a base and trimethylsulfoxonium iodide and add the mixture tothe compound (21). Examples of the inorganic base as used herein are notparticularly limited, but include, sodium hydride, potassium hydroxide,sodium carbonate, potassium carbonate, sodium bicarbonate, potassiumphosphate, caesium carbonate, and the like.

(Starting Material Synthesis 6)

A compound (11) can be obtained by subjecting a compound (6) to reactionwith silver cyanate. This reaction can be carried out using similarconditions as for the reaction of a compound (15) with silver cyanate in(Production Process 1), except for use of excess tert-butanol in thepresence of triethylamine, instead of excess NH₃, in the cyclizationstep.

(Starting Material Synthesis 7)

A compound (23) can be obtained by subjecting a compound (3) to reactionwith nucleophilic methylation reagent and dehydration reaction.

In this reaction, a mixture of the compound (3) and nucleophilicmethylation reagent is stirred under any temperature condition fromcooling to heating and refluxing, and preferably at −78° C. to 80° C.,usually for 0.1 hours to 5 days, in a solvent which is inert to thereaction or without a solvent. Examples of the solvent as used hereinare not particularly limited, but include aromatic hydrocarbons such asbenzene, toluene, xylene, and the like, ethers such as diethyl ether,tetrahydrofuran, dioxane, dimethoxyethane, and the like and a mixturethereof. Examples of the nucleophilic methylation reagent as used hereinare not particularly limited, but include methylmagnesium bromide,methylmagnesium chloride, methylmagnesium iodide, methyllithium, and thelike.

After the methylation step, an acid is added to the reaction mixture.And, the reaction mixture is stirred under any temperature conditionfrom cooling to heating and refluxing, and preferably at 0° C. to 80°C., usually for 0.1 hours to 5 days. Examples of the organic acid asused herein are not particularly limited, but include, acetic acid,trifluoroacetic acid, p-toluenesulfonic acid, methanesulfonic acid andthe like, examples of the inorganic acid as used herein are notparticularly limited, but include, hydrochloric acid, sulfuric acid,potassium hydrogen sulfate and the like.

A compound (24) can be obtained by subjecting a compound (23) to acyclization reaction.

First, a mixture of the compound (23) and a sulfonyl halide is stirredunder any temperature condition from cooling to heating and refluxing,and preferably at −18° C. to 50° C., usually for 0.1 hours to 5 days, ina solvent which is inert to the reaction or without a solvent in thepresence of a base. Examples of the sulfonyl halide as used herein arenot particularly limited, but include methanesulfonyl chloride, tosylchloride, and the like. Examples of the solvent as used herein are notparticularly limited, but include aromatic hydrocarbons such as benzene,toluene, xylene, and the like, ethers such as diethyl ether,tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenatedhydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform,and the like, ethyl acetate, acetonitrile, and a mixture thereof.Examples of the organic base as used herein are not particularlylimited, but include pyridine, triethylamine, N,N-diisopropylethylamine,N-methylmorpholine, methyllithium, n-butyllithium and the like, or aninorganic base such as potassium carbonate, sodiumbis(trimethylsilyl)amide, sodium carbonate, potassium hydroxide, caesiumcarbonate, sodium hydroxide, sodium hydride and the like. In some cases,compound (23) can be pretreated with the base for the smooth progress ofthe reaction before addition of the sulfonyl halide.

After the sulfonylation step, a base is added to the reaction mixture.And, the reaction mixture is stirred under any temperature conditionfrom cooling to heating and refluxing, and preferably at 0° C. to 80°C., usually for 0.1 hours to 5 days. Examples of an organic base as usedherein are not particularly limited, but include pyridine,triethylamine, N,N-diisopropylethylamine, N-methylmorpholine,methyllithium, n-butyllithium and the like, or an inorganic base such aspotassium carbonate, potassium bis(trimethylsilyl)amide, sodiumcarbonate, potassium hydroxide, caesium carbonate, sodium hydroxide,sodium hydride and the like.

The compounds of the formula (I) can be isolated and purified as theirfree compounds, salts, hydrates, solvates, or polymorphic crystalsubstances thereof. The salts of the compound of the formula (I) can beprepared by carrying out a conventional salt forming reaction.

Isolation and purification are carried out by employing ordinarychemical operations such as extraction, fractional crystallization,various types of fractional chromatography, and the like.

Various isomers can be prepared by selecting an appropriate startingcompound or separated by using the difference in the physicochemicalproperties among the isomers. For example, the optical isomers can beobtained by means of a general method for designing optical resolutionof racemic products (for example, fractional crystallization forinducing diastereomer salts with optically active bases or acids,chromatography using a chiral column or the like, and others), andfurther, the isomers can also be prepared from an appropriate opticallyactive starting compound.

The pharmacological activity of the compounds of the formula (I) wasconfirmed by the tests shown below.

TEST EXAMPLE Inhibition of Beta-Secretase Activity Test Example 1Measurement of BACE1 Inhibition by Fluorescence Resonance EnergyTransfer (FRET)

Potency of test compounds were determined by measurement of theirinhibition of BACE1 activity toward a fluorescent substrate. Experimentswere performed by reference to the procedure as described in Ermolieff,et al. (Biochemistry 39:12450-12456 (2000), the teachings of which areincorporated hereby in their entirety). Briefly, the recombinantprotease unit of BACE1 was prepared from E. coli expression as inclusionbodies, refolded, and purified as described in Lin, et al., (Proc. Nat.Acad. Sci. 97:1456-1460 (2000)). Fluorogenic substrate,MCA-SEVNLDAEFK(DNP)—NH₂ (SEQ ID NO:1) was purchased. (M−2485, BachemAmericas, Torrance, Calif.). The substrate was derived from 10 aminoacids of the human amyloid precursor protein (APP), with the Swedishvariant amino acids at the beta-secretase cleavage site. The terminalamino acid was modified from arginine to lysine to facilitatederivatization with a functional group for detection byautofluorescence. The amino acid sequence of the “core” peptide of thesubstrate is SEVNLDAEFK (SEQ ID NO:2). The amino terminus wasderivatized with (7-methoxycoumarin-4-yl)acetyl (MCA), and the epsilonamine of the lysine side chain of the terminal residue (K in sequenceSEVNLDAEFK (SEQ ID NO:2)) was derivatized with 2,4-dinitrophenyl (DNP).Assays were performed in a buffer of 0.1 M sodium acetate, pH 4.4, 0.08%3-[(3-Cholamidopropyl)dimethylammonio]propanesulfonate (CHAPS), 0.005%Tween80. BACE1 enzyme (final concentration 65 nM) was pre-incubated withtest compounds for 15 minutes at room temperature. Fluorescenceintensities were measured 60 minutes after addition of the substrate(final concentration 3 μM) by Tecan Safire2™. An excitation wavelengthof 328 nm and an emission wavelength of 393 nm were used. For thecalculation of % inhibition, fluorescence intensity without compoundswas defined as the value for 0% inhibition and fluorescence intensitywithout the enzyme was defined as the value for 100% inhibition. Thevalues of IC₅₀ were calculated by GraphPad Prism version 5.

Moreover, the inhibition constants, Ki, were determined as described inErmolieff, et al. (Biochemistry 39:12450-12456 (2000)). Briefly, thehydrolysis of the fluorogenic substrate, for a series of mixtures withconstant enzyme, but increasing inhibitor concentration was carried outin the same manner as described in the method for Test Example 1.Quantification of enzymes was achieved by active-site titration using atight-binding inhibitor. The inhibition constant, Ki, was determinedfrom plot of activity vs. inhibitor concentration based on the equationdescribed in Ermolieff, et al. (Biochemistry 39:12450-12456 (2000)).

The results of the representative compounds are shown in [Table. 1]below.

The inhibition constants Ki of Example 89 and Example 98 compoundsdescribed in Pamphlet of International Publication WO2011/123674 weredetermined. In the result, the Ki value of Example 89 compound was 0.241μM, and the Ki value of Example 98 compound was 4.087 μM.

Herein, the structure of Example 89 compound described in Pamphlet ofInternational Publication WO2011/123674 is

and this compound is a racemic mixture.

The structure of Example 98 compound described in Pamphlet ofInternational Publication WO2011/123674 is

and this compound is a racemic mixture.

The results of the representative compounds are shown in [Table. 1]below.

Test Example 2 Measurement of BACE1 Inhibition by Time ResolvedFluorescence Resonance Energy Transfer (TR-FRET)

Potency of compounds were also measured using another fluorogenicsubstrate, TruPoint BACE1 Substrate Eu-CEVNLDAEFK-QSY 7 (SEQ ID NO:3)(AD0258, PerkinElmer, Boston Mass.). This substrate also has Swedishvariant amino acids at the β-secretase cleavage site, with a fluorescenteuropium (Eu) chelate coupled to one end and a quencher of europiumfluorescence (QSY7) coupled to the other end via lysine. If the samplecontains BACE1 activity, the Eu chelate and the quencher will beseparated as the substrate is cleaved. The Eu-signal increases and itcan be measured by time-resolved fluorometry, EnVision™, 30 minutesafter the substrate (final concentration 300 nM) was added.

The experiment was basically conducted in a way similar to Test Example1 above.

The results of the representative compounds are shown in [Table. 1]below.

Test Example 3 Measurement of Aβ Production Inhibition in Cell

The potency of compounds against BACE1 activity was determined in acellular assay of Aβ production. Human SK-N-BE(2) neuroblastoma cells(ATCC No. CRL-2271) were plated at 96,000 cells/well/100 μL in 96-wellplates in RPMI1640 medium/10% fetal bovine serum(FBS)/penicillin-streptomycin and cultured for 24 hours at 37° C., 5%CO₂. Test compounds were dissolved in dimethyl sulfoxide and dilutedwith dimethyl sulfoxide and put into RPMI1640/5%FBS/penicillin-streptomycin media (final dimethyl sulfoxideconcentration is 0.5%). The culture media in 96-well plates werereplaced by 125 μL/well of the media containing test compounds. Afterincubation for 6 hours at 37° C., 5% CO₂, 30 μL of the media weretransferred into a fresh 96-well plate and used for Aβ40 assay by anenzyme-linked immunosorbent assay (ELISA) kit (#27718, Immuno-BiologicalLaboratories, Japan) according to the manufacturer's protocol. Cellviability was measured by CellTiter-Glo™ Luminescent Cell ViabilityAssay (#7571, Promega) after removal of 30 μL of the media for the Aβassay. CellTiter-Glo Substrate was dissolved into CellTiter-Glo Bufferand added to the plates in 95 μL/well. After shaking the plates for 2minutes, the whole sample was transferred into a white 96-well plate andluminescence was measured for ATP quantification as the cell viability.Aβ concentration measured by ELISA was normalized by the viability ofthe corresponding cells. The values of IC₅₀ were calculated by GraphPadPrism version 5.

The results of the representative compounds are shown in [Table. 1]below.

Test Example 4 Brain Aβ Reduction in Rats

Effects on brain Aβ reduction in rats were determined with reference tothe method described in the WO2012/054510. It was confirmed that some ofthe compounds of the formula (I) exhibit brain Aβ reduction in rats.Concretely, the test was conducted with the method as below.

Formulation

Test compounds were prepared in a vehicle of 35% HPβCD in H₂O. The testcompound was formulated the same day as oral dosing. Doses (see [Table.1]) were based on the free base equivalent. Sonication was used whererequired to facilitate the formulation.

Test Species

Male Sprague-Dawley rats (150-300 grams) were obtained from CharlesRiver Japan (Atsugi, Japan) and were given approximately 4 days ofacclimation. Food and water was made available ad libitum throughout thestudy. Animals were visually inspected for health before being includedinto the study group, and were randomly assigned to the treatment andcontrol groups to achieve similar group mean body weights. The dosingsolution (dose volume 5 mL/kg) was administered directly into thestomach using a rodent gavage needle. Control animals received oraladministration of equivalent volume of the vehicle.

Sampling Methods

At a time post-dose (e.g., 3 hours; see [Table. 1]), animals wereeuthanized with isoflurane. Blood was collected from the inferior venacava using syringe flushed with EDTA 2K and placed on ice. Plasma wasseparated using centrifugation at 15,000 rpm (20,400×g) for 5 minutes at4° C. and subsequently stored at −80° C. After blood sampling, CSF(cerebrospinal fluid) was carefully withdrawn from the cisterna magnausing a 29 gauge needle after a quick dissection to expose theatlantooccipital membrane. The CSF samples were centrifuged at 15,000rpm (20,400×g) for 5 minutes to confirm free of blood contamination andstored at −80° C. Immediately after decapitation, the hippocampus wereisolated on ice, and quickly frozen in liquid nitrogen and stored at−80° C.

Extraction of Brain Aβ42

Fragments of the hippocampus were weighed while frozen. A 10-fold volume(w:v) of TBS (Tris-buffered saline) supplemented with a Complete Miniprotease inhibitor tablet (catalog number: 11 836 153 001, RocheDiagnostics, In, USA) was added. The hippocampus were homogenized usingsonication on ice in microcentrifuge tube. Resulting homogenates werecentrifuged at 100,000×g for 1 hour in a refrigerated centrifuge at 4°C. Supernatants were collected as soluble fraction.

Determination of Aβ42

Concentration of Aβ42 in the extract of hippocampus, plasma, and CSFwere analyzed using ELISA (Human/Rat Aβ42 ELISA, catalog number292-64501, Wako Pure Chemical Industries, Ltd. Japan). Eachconcentration of Aβ42 was divided by the mean concentration of Aβ42 ofvehicle-treated group, and these ratios were converted to percentages.

The results of the representative compounds are shown in [Table. 1]below.

Test Example 5 hERG (Human Ether-a-go-go Related Gene) Analysis

hERG Inhibition

The hERG potassium current was measured in a hERG-stably-expressingChinese hamster ovary K1 (CHO) cells. The experiments were performedusing an automated planar patch-clamp system QPatch HTX (SophionBioscience A/S). The application of pressure for forming gigaseals andwhole-cell patch clamp configuration were established using the QPatchassay software. Patch-clamp experiments were performed in voltage-clampmode and whole-cell currents were recorded. The following stimulationprotocol was applied to investigate the effects of compounds on hERGpotassium channel.

The membrane potential was held at −80 mV and repetitively (every 15seconds) depolarized to +20 mV for 4800 milliseconds after the pulse to−50 mV for 20 milliseconds served to define the baseline, followed byrepolarizing step to −50 mV for 5000 milliseconds to evaluate of thetail current amplitude. Experiments were conducted at room temperature.

Effects of compounds were determined from cumulative applications ofincreasing 6 concentrations and calculated as percent of blockedcurrent. The data points were fitted with Hill equation to calculatehalf-maximal inhibition concentrations (IC₅₀). The maximum compoundconcentration tested in the assay was 10 μM for some compounds. If lessthan 50% inhibition was achieved at the 10 μM compound concentration,the IC₅₀ is reported as >10 μM.

The test solution includes:

Extracellular solution: 2 mM of CaCl₂, 1 mM of MgCl₂, 10 mM of HEPES, 4mM of KCl, 145 mM of NaCl, and 10 mM of glucose; and pH adjusted to 7.4with NaOH,

Intracellular solution: 5.374 mM of CaCl₂, 1.75 mM of MgCl₂, 10 mM ofHEPES, 10 mM of EGTA, 120 mM of KCl, and 4 mM of ATP, and pH adjusted to7.2 with KOH.

hERG selectivity

Selectivity of BACE1 inhibition over hERG inhibition was calculated bydividing hERG IC₅₀ by BACE1 Ki. The results of the representativecompounds are shown in [Table. 1] below. As mentioned above, someresults from the hERG assay are necessarily reported as >10 μM. Usingthese values in the calculation of selectivity will necessarily causethe selectivity values to be characterized as “>” or “greater than” thecalculated ratio.

The hERG IC₅₀ values of Example 89 and Example 98 compounds described inPamphlet of International Publication WO2011/123674 were determined. Inthe result, the IC₅₀ value of Example 89 compound was 0.44 μM, and theIC₅₀ value of Example 98 compound was 9.46 μM. Moreover, selectivityvalues of BACE1 inhibition over hERG inhibition of these compounds werecalculated. In the result, the selectivity value of Example 89 compoundwas 1.8, and the selectivity value of Example 98 compound was 2.3.

It is considered to be desirable that compounds show their primarypharmacological effect with selectivity over hERG inhibition (Jamiesonet al. J. Med. Chem. 2006, 49, 5029). Compounds with lower hERGselectivity are considered to have higher risk to cause QTcprolongation, which may eventually lead to drug-induced arrhythmia andsudden deaths. For example, hERG selectivity around or less than 10-foldover hERG inhibition are recognized to have a concern for high risk ofQTc prolongation (Kongsamut et al. Eur. J. Pharmacol. 2002, 450, 37. andMinotti, Cardiotoxicity of Non-Cardiovascular Drugs, Wiley, 2010. p.65), whereas compounds with selectivity around or larger than 100-foldare recognized to be more favorable (Pajouhesh et al. Bioorg. Med. Chem.Lett. 2012, 22, 4153; Micheli et al. J. Med. Chem. 2010, 53, 374.).

The results of the representative compounds are shown in [Table. 1]below.

In [Table. 1], Ex means Example Number, and “Test Example X” refers tothe protocol described above used to obtain the data. Moreover, RPprefixed before the numeral shows the compound of Reference Example.

TABLE 1 Test Test Test Test Test Test Test Example 1 Example 1 Example 2Example 3 Example 4 Example 5 Example 5 IC₅₀ BACE1 MCA IC₅₀ IC₅₀ Aβ42hERG hERG Ex. (μM) Ki (μM) (μM) (μM) reduction (%) IC₅₀ (μM) selectivityRP 73.3 1a RP 138 1b RP 59.6 2 RP 33.8 3 RP 12.6 7.2 >10 4 RP 1.560.59 >10 5 RP 11.7 3.9 >10 6 RP 39.0 7a RP 39.7 7b RP 17.3 >10 9a RP24.4 >10 9b RP 61.3 8 RP 7.21 >10 10 RP 15.6 11a RP 17.4 11b RP 38.7 12aRP 81.3 12b RP 11.1 13a RP 19.3 13b RP 19.0 14a RP 44.2 14b RP 1.721.1 >10 15a RP 1.01 0.44 7.93 15b RP 9.54 >10 16a RP 4.57 4.8 >10 16b RP30.2 >10 17 RP 41.3 18 RP 2.31 0.43 19 RP 2.71 0.58 20 RP 1.35 0.41 21RP 2.60 2.9 22 RP 0.519 0.90 23 RP 37.9 32.9 24 RP 36.9 25 RP 3.75 0.06926 27 0.0822 0.0382 0.0094 >10 >262 28 0.154 0.117 0.019 >10 >86 RP0.276 0.240 0.022 >10 >42 29 30 0.0975 0.0515 0.0023 >10 >194 RP 1.061.03 0.061 >10 >10 31 RP 0.418 0.378 0.022 2.17 6 32 RP 1.52 0.95 33 RP1.24 0.26 >10 34 RP 26.0 35 RP 2.90 0.81 36 RP 55.4 37 RP 17.3 38 RP46.9 40 RP 24.8 >30 41 RP 16.1 5.9 >10 42 RP 1.96 1.4 >10 43 RP 0.6020.098 44 RP 1.04 0.057 >10 45 RP 8.94 1.4 >10 46 RP 2.33 1.4 47 RP 1.130.69 48 RP 48.6 >10 51a RP 0.172 0.137 0.072 >10 51b RP 0.161 0.075 4.8352a RP 0.157 0.123 0.099 >10 53b RP 0.406 0.079 >10 54 RP 0.74 >10 56 RP3.69 0.61 3.48 57 RP 1.48 0.18 5.81 58 RP 1.11 0.076 6.9 59 RP 1.48 0.162.78 60 RP 0.35 9.22 61 RP 1.33 1.0 4.43 62 RP 7.90 3.3 4.48 63 RP 6.122.3 1.67 64 RP 0.388 0.049 9.77 65 RP 135 >10 66 RP 96.5 >10 67 RP 0.3450.50 6.05 68 RP 0.397 0.040 >10 69 RP 1.75 0.96 >10 70 RP 1.08 0.11 >1071 RP 0.817 0.068 8.22 72 RP 0.827 0.070 73 RP 35.4 74 RP 3.67 1.1 >1075 RP 13.4 >10 76 RP 1.43 0.92 >10 77 RP 2.05 1.3 >10 78 RP 1.690.45 >10 79 RP 1.91 0.71 >10 80 RP 1.89 0.51 >10 81 RP 0.348 0.036 >1082 RP 0.285 0.255 0.047 >10 83 RP 0.735 0.41 >10 84 RP 0.319 0.077 >1085 RP 0.143 0.107 0.058 7.94 86 RP 1.55 0.43 87 RP 2.51 1.4 >10 88 RP4.67 >10 89 RP 0.901 0.50 >10 90 RP 0.656 0.34 >10 91 RP 0.124 0.08790.037 5.34 92 RP 0.629 0.37 >10 93 RP 0.862 0.38 >10 94 RP 0.7400.41 >10 95 RP 0.126 0.868 0.022 >1 96 RP 20.2 2.6 >10 97 RP 64.1 >10 98RP 2.97 1.9 >10 99 RP 2.62 0.25 >10 100 RP 7.65 5.28 101 RP 1.610.24 >10 102 RP 2.19 0.45 >10 103 RP 1.13 0.30 3.25 104 RP 2.94 0.34 7.9105 RP 1.83 0.16 5.75 106 RP 1.06 0.46 2.91 107 RP 0.383 0.354 0.3160.096 9.4 108 RP 0.152 0.120 0.048 >1 109 RP 0.137 0.0904 0.0092 >1 110RP 0.287 0.252 0.14 1.98 111 RP 0.400 0.66 4.32 112 RP 1.11 0.47 113 RP68.3 114 RP 3.31 1.1 >10 115 RP 1.27 0.70 >10 116 RP 4.40 3.0 >10 117 RP1.81 0.99 >10 118 RP 3.56 119 RP 1.89 2.3 120 RP 3.37 121 RP 2.74 1.1122 RP 3.39 123 RP 3.59 124 RP 2.31 0.58 125 RP 0.696 1.2 126 RP 2.271.8 127 RP 1.68 1.5 128 RP 2.21 7.3 129 RP 1.87 1.7 130 RP 2.02 3.2 131RP 2.45 1.2 132 RP 1.67 2.8 133 RP 3.63 134 RP 3.27 1.8 135 RP 0.6350.66 136 RP 0.773 0.83 137 RP 1.67 1.5 138 RP 0.252 0.215 0.14 139 RP0.389 0.41 140 RP 0.903 1.30 141 RP 0.401 0.15 >1 142 RP 1.26 143 RP0.879 0.46 144 RP 0.933 145 RP 0.226 0.188 0.21 146 RP 0.621 0.17 147 RP0.614 0.48 148 RP 1.593 1.1 149 RP 0.759 0.45 150 RP 1.36 0.28 151 RP0.688 0.11 152 RP 1.13 0.13 153 RP 0.313 0.032 >1 154 RP 0.786 0.63 155RP 2.61 0.18 156 RP 2.80 0.66 157 RP 0.930 0.37 158 RP 2.12 159 RP 0.981160 RP 1.41 161 RP 1.04 162 RP 1.064 163 RP 2.314 164 RP 0.760 165 RP3.63 166 RP 0.869 167 RP 0.760 0.31 168 RP 1.41 0.859 0.90 169 RP 1.000.588 0.40 170 RP 2.24 3.04 3.1 171 RP 2.24 2.16 172 RP 0.401 0.2390.073 173 RP 3.35 2.18 3.4 174 RP 3.23 1.55 175 RP 0.994 0.85 176 RP2.12 1.5 177 RP 1.15 0.747 0.81 178 RP 3.24 2.06 1.4 179 RP 2.03 1.770.49 180 RP 2.59 2.13 0.82 181 RP 1.45 2.5 182 RP 1.08 0.880 183 RP 2.822.72 184 RP 2.05 1.27 0.76 185 RP 2.42 186 RP 3.16 1.92 187 RP 50.6 >10188 RP 0.661 0.623 0.043 >10 >16 189 190 0.125 0.0835 0.0041 >10 >120 RP0.248 0.215 0.027 >10 >47 191 RP 0.318 0.280 0.030 >10 >36 192 RP 1.040.081 >10 >120 193 RP 0.349 0.313 0.028 4.91 16 194 195 0.109 0.06450.0032 >10 >155 RP 0.380 0.342 0.053 3.65 11 196 197 0.118 0.07200.0057 >10 >139 RP 14.9 >1 198 RP 0.736 0.24 4.25 199 RP 0.128 0.09210.026 >10 200 RP 0.113 0.0711 0.0087 201 RP 0.145 0.113 0.026 7.73 202RP 0.109 0.0676 0.0049 203 RP 0.388 0.24 >10 204 RP 0.126 0.0825 0.0225.22 205 RP 4.91 >30 >10 206 RP 0.460 0.16 >10 207 RP 7.76 6.1 >10 208RP 2.00 0.83 >10 209 RP 4.95 3.7 210 RP 19.4 211 RP 17.6 >10 212 RP 39.66.3 >10 213 RP 40.3 214 RP 1.21 0.24 >10 215 RP 55.8 216 218 0.04470.0103 0.00078 24% 20.67 2007 (3 mg/kg, 2.5 hours) 31% (10 mg/kg, 2.5hours) 219 0.0444 0.0102 0.0418 0.0022 >10 >980 220 0.0765 0.0429 0.003121% >10 >233 (3 mg/kg, 1.5 hours) 27% (10 mg/kg, 1.5 hours) 221 0.08180.0378 0.00016 24% 3.25 86 (1 mg/kg, 3 hours) 42% (3 mg/kg, 3 hours) 2220.138 0.0915 0.00062 >10 >109 223 0.0991 0.0612 0.0031 19% 20.77 339 (1mg/kg, 3 hours) 34% (3 mg/kg, 3 hours) 224 0.0914 0.0527 0.00063 21%15.1 287 (1 mg/kg, 3 hours) 35% (3 mg/kg, 3 hours) 227 0.107 0.07920.0080 >10 >126 228a 30.3 30.2 >10 >0.3 229a 47.6% inhibition at 30microM

As described above, it was confirmed that the representative compoundsof the formula (I) have β-secretase inhibitory activities, Aβ productioninhibitory activities, and Aβ reduction activities and can be thereforeused for diseases or conditions associated with and/or mediated byβ-secretase activity, hydrolysis of a β-secretase cleavage site of anamyloid precursor protein, and/or β-amyloid protein accumulation, suchas Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease,especially, Alzheimer's disease, or the like.

Determination of Absolute Stereochemistry by Vibrational CircularDichroism (VCD) Spectrometry Measurements

The infrared and VCD spectra were recorded on a Bio tools ChiralIR-2X™Vibrational Circular Dichroism (VCD) spectrometer.

The infrared and VCD spectra were measured in CDCl₃ solution placed in a100 μm path length cell with BaF₂ windows at 4 cm⁻¹ resolution and theirdata collection was performed for 5 hours.

Calculations

Conformational searches were executed by using CONFLEX™ ver.6 program.

The geometry optimizations and the calculations of theoretical infraredand VCD spectra were implemented using density functional theory withB3LYP functional and 6-31G(d) basis set on Gaussian 09.

By comparison of measured and calculated spectra, absolutestereochemistry of Ex. 228a and 228b, Ex. 229a and 229b, Ex. 225a and225b compounds were assigned. Based on the absolute stereochemistry ofEx. 228b and Ex. 229b compound, the absolute stereochemistry of theirprecursor, Reference Example 226 compound was determined.

Powder X-Ray Diffraction

The powder X-ray diffraction was measured using RIGAKU RINT-TTRIIdiffractometer under the conditions of a tube: Cu, a tube current: 300mA, a tube voltage, 50 kV, a sampling width: 0.02°, a scanning speed:4°/minute, a wavelength: 1.54056 angstroms, and a measurementdiffraction angle (28): 2.5 to 40°.

Furthermore, the term “about” in the characteristic peaks of powderX-ray diffraction shown at angles 2θ denotes ±0.2°, in anotherembodiment, ±0.1°. Each crystal can be characterized by a powder X-raydiffraction spectrum, but with the powder X-ray diffraction, crystallattice intervals and overall patterns are important for identificationof crystals in terms of the properties of the data, and since therelative intensity may vary slightly depending on the direction ofcrystal growth, the particle size, and the measurement conditions, itshould not be strictly construed.

Formulations

The pharmaceutical composition containing one or two or more kinds ofthe compound represented by the formula (I) or salts thereof as anactive ingredient can be prepared using excipients that are usually usedin the art, that is, excipients for pharmaceutical preparation, carriersfor pharmaceutical preparation, and the like.

A “pharmaceutically acceptable carrier” refers to an ingredient in apharmaceutical composition, other than an active ingredient, which isnontoxic to a subject. A pharmaceutically acceptable carrier includes,but is not limited to, a buffer, excipient, stabilizer, or preservative.

Administration can be accomplished either by oral administration viatablets, pills, capsules, granules, powders, solutions, and the like, orparenteral administration, such as intraarticular, intravenous, orintramuscular injections, and the like, suppositories, ophthalmicsolutions, eye ointments, transdermal liquid preparations, ointments,transdermal patches, transmucosal liquid preparations, transmucosalpatches, inhalers, and the like.

The solid composition for use in the oral administration according tothe present invention is used in the form of tablets, powders, granules,or the like. In such a solid composition, one or more activeingredient(s) are mixed with at least one inactive excipient. Accordingto a conventional method, the composition may contain inactiveadditives, such as a lubricant such as magnesium stearate, adisintegrating agent such as sodium carboxymethyl starch and the like, astabilizer, or a solubilization assisting agent. If necessary, tabletsor pills may be coated with sugar or a film of a gastric or entericcoating substance.

The liquid composition for oral administration contains pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, elixirs, or thelike, and also contains generally used inert diluents, for example,purified water or ethanol. In addition to the inert diluent, the liquidcomposition may also contain auxiliary agents, such as a solubilizationassisting agent, a moistening agent, and a suspending agent, sweeteners,flavors, aromatics, and antiseptics.

The injections for parenteral administration include sterile aqueous ornon-aqueous solution preparations, suspensions and emulsions. Theaqueous solvent includes, for example, distilled water for injection andphysiological saline. Examples of the non-aqueous solvent includealcohols such as ethanol. Such a composition may further contain atonicity agent, an antiseptic, a moistening agent, an emulsifying agent,a dispersing agent, a stabilizing agent, or a solubilizing aid. Theseare sterilized, for example, by filtration through a bacteria retainingfilter, blending of a bactericide, or irradiation. In addition, thesecan also be used by preparing a sterile solid composition, anddissolving or suspending it in sterile water or a sterile solvent forinjection prior to its use.

The agent for external use includes ointments, plasters, creams,jellies, poultices, sprays, lotions, eye drops, eye ointments, and thelike. The agents contain generally used ointment bases, lotion bases,aqueous or non-aqueous liquid preparations, suspensions, emulsions, andthe like.

As the transmucosal agents such as an inhaler, a transnasal agent, andthe like, those in the form of a solid, liquid, or semi-solid state areused, and can be prepared in accordance with a conventionally knownmethod. For example, a known excipient, and also a pH adjusting agent,an antiseptic, a surfactant, a lubricant, a stabilizing agent, athickening agent, or the like may be appropriately added thereto. Fortheir administration, an appropriate device for inhalation or blowingcan be used. For example, a compound may be administered alone or as apowder of formulated mixture, or as a solution or suspension incombination with a pharmaceutically acceptable carrier, using aconventionally known device or sprayer, such as a measuredadministration inhalation device, and the like. A dry powder inhaler orthe like may be for single or multiple administration use, and a drypowder or a powder-containing capsule may be used. Alternatively, thismay be in a form such as a pressurized aerosol spray which uses anappropriate ejection agent, for example, a suitable gas such aschlorofluoroalkane, hydrofluoroalkane, carbon dioxide, and the like, orother forms.

In oral administration, the daily dose is generally from about 0.001 to100 mg/kg, preferably from 0.1 to 30 mg/kg, and more preferably 0.1 to10 mg/kg, per body weight, administered in one portion or in 2 to 4divided portions. In the case of intravenous administration, the dailydose is suitably administered from about 0.0001 to 10 mg/kg per bodyweight, once a day or two or more times a day. In addition, atransmucosal agent is administered at a dose from about 0.001 to 100mg/kg per body weight, once a day or two or more times a day. The doseis appropriately decided in response to the individual case by takingthe symptoms, the age, and the gender, and the like into consideration.

The compound of the formula (I) can be used in combination with varioustherapeutic or prophylactic agents for the diseases, in which thecompound of the formula (I) is considered effective, as described above.The combined preparation may be administered simultaneously orseparately and continuously, or at a desired time interval. Thepreparations to be co-administered may be a blend or preparedindividually.

EXAMPLES

Hereinbelow, the preparation methods for the compound of the formula (I)will be described in more detail with reference to Examples. Further,the present invention is not limited to the preparation methodsdescribed in the specific Examples, Reference Examples and PreparationExamples as described below, but the compound of the formula (I) can beprepared by any combination of the preparation methods or the methodsthat are apparent to a skilled person in the art, particularly in viewof the detailed teachings provided herein.

Furthermore, the following symbols are used in the Examples, ReferenceExamples, Preparation Examples, and Tables as described below.

Rf: Preparation Example Number,

RP: Reference Example Number,

Ex: Example Number,

No.: Compound No.,

Data: Physicochemical data,

ESI+: representing m/z values in ESI-MS (positive ions), andrepresenting [M+H]⁺ peaks unless otherwise specified,

APCI/ESI+: m/z value in APCI/ESI-MS (positive ions), and representing[M+H]⁺ peaks unless otherwise specified,

EI: representing m/z values in EI-MS (positive ions), and representing[M]⁺ peaks unless otherwise specified,

CI+: representing m/z values in CI-MS (positive ions), and representing[M+H]⁺ peaks unless otherwise specified,

NMR-DMSO-d₆: δ (ppm) in ¹H-NMR in DMSO-d₆,

NMR-CDCl₃: δ (ppm) in ¹H-NMR in CDCl₃,

Structure: Structural formula (In case HCl is described in a structuralformula, a compound represented by a structural formula forms a saltwith HCl. Compounds having a double bond described by a cross linerepresents mixtures of a cis compound and a trans-compound),

rel-: representing relative configuration,

Syn: Preparation method (in which E prefixed before the numeral showsthat the compound is prepared by the similar preparation method as thecompound having the Example Number, R prefixed before the numeral showsthat the compound is prepared by the similar preparation method as thecompound having the Preparation Example Number and RP prefixed beforethe numeral shows that the compound is prepared by the similarpreparation method as the compound having the Reference Example Number),

Boc/BOC: tert-butoxycarbonyl,

CHCl₃: chloroform,

CH₂Cl₂: dichloromethane,

CO₂: carbon dioxide,

Cs₂CO₃: caesium carbonate,

CuBr: copper (I) bromide,

CuI: copper (I) iodide,

DAST: N,N-diethylaminosulfur trifluoride,

DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene,

DIBAL-H: diisobutylaluminium hydride,

DMAP: N,N-dimethyl-4-aminopyridine,

DMF: N,N-dimethylformamide,

DMSO: dimethyl sulfoxide,

Et₃N: triethylamine,

AcOEt/EtOAc: ethyl acetate,

EtOH: ethanol,

Et₂O: diethyl ether,

HCOOH: formic acid,

HCl: hydrogen chloride,

H₂O: hydrogen oxide,

HPLC: high performance liquid chromatography,

IPE, iPr₂O: diisopropyl ether,

K₂CO₃: potassium carbonate,

K₃PO₄: potassium phosphate,

LiBH₄: lithium borohydride,

MeCN: acetonitrile,

MsCl: methanesulfonyl chloride,

MeMgBr: methylmagnesium bromide,

MeOH: methanol,

MgSO₄: anhydrous magnesium sulfate,

n-BuLi: n-butyllithium,

NMP: 1-methyl-2-pyrrolidone,

NaOH: sodium hydroxide,

NaHCO₃: sodium hydrogen carbonate,

Na₂CO₃: sodium carbonate,

Na₂S₂O₃: sodium thio sulfate,

Na₂SO₄: anhydrous sodium sulfate,

Na₂SO₄.10H₂O: sodium sulfate decahydrate,

NH₄Cl: ammonium chloride,

PdCl₂(dppf): [1,1′-Bis(diphenylphosphino)ferrocene]palladium(II)dichloride,

Pd(OAc)₂: palladium(II) acetate,

Pd(PPh₃)₄: tetrakis(triphenylphosphine)palladium(0),

PdCl₂(PPh₃)₂: bis(triphenylphosphine)palladium (II) chloride,

PPh₃: triphenylphosphine,

PtO₂: platinum (IV) oxide,

SiO₂: silicon dioxide,

THF: tetrahydrofuran,

TsOH.H₂O: p-toluenesulfonic acid monohydrate,

TMSOTf: Trimethylsilyl trifluoromethanesulfonate.

Preparation Example 1

To a mixture of 6-bromo-4-methylene-4H-spiro[chromene-3,3′-oxetane] (351mg, 1.31 mmol), silver cyanate (295 mg, 1.97 mmol), EtOAc (1.7 mL), andMeCN (3.5 mL) was added a mixture of iodine (500 mg, 1.97 mmol) andEtOAc (5.3 mL) in an ice-water bath. After stirring for 1.5 hours at thesame temperature, the mixture was stirred for 30 minutes at ambienttemperature. The mixture was filtered through celite pad (washed withEtOAc), and the filtrate was washed with saturated aqueous Na₂S₂O₃ andbrine, dried over MgSO₄ and filtered. After concentration of thefiltrate at reduced pressure, tert-butyl alcohol (4.4 mL) andtriethylamine (0.183 mL, 1.31 mmol) were added to the residue, and themixture was stirred overnight under reflux. The reaction mixture wascooled down to ambient temperature, concentrated at reduced pressure togive crude6′-bromo-2H-dispiro[1,3-oxazolidine-4,4′-chromene-3′,3″-oxetan]-2-one.

Preparation Example 2

A mixture of di-tert-butyl

[6′-(3-methoxyprop-1-yn-1-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(34.7 mg, 0.067 mmol) and 10% palladium on carbon (7 mg) in EtOH (1.4mL) was stirred for 13 hours under a hydrogen atmosphere (4.5 kgf/cm²).The mixture was filtered off, and the filtrate was evaporated to givecrude di-tert-butyl[6′-(3-methoxypropyl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(34.7 mg).

Preparation Example 8

To a solution of6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(643 mg, 1.82 mmol) in THF (12.9 mL) were added 4-dimethylaminopyridine(11 mg, 0.091 mmol) and di-tert-butyl dicarbonate (1.19 g, 5.46 mmol).The mixture was stirred overnight at ambient temperature. The reactionmixture was evaporated off at reduced pressure. The residue was purifiedwith column chromatography on silica gel (hexane-EtOAc, a lineargradient of EtOAc from 0 to 20%) to give di-tert-butyl(6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(890 mg).

Preparation Example 22

The mixture of6′-bromodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(1.1 g, 3.4 mmol), di-tert-butyl dicarbonate (2.2 g, 10 mmol), andN,N-dimethylpyridin-4-amine (21 mg, 0.17 mmol) in THF (21 ml) wasstirred for 3 hours at ambient temperature and for 5 hours at 50° C. Themixture was concentrated in vacuo. The residue was purified by silicagel chromatography (hexane:EtOAc=100:0-80:20) to give di-tert-butyl(6′-bromodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl)imidodicarbonate(1.5 g).

Preparation Example 23

The mixture of6′-bromodispiro[cyclobutane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(185 mg, 0.57 mmol), di-tert-butyl dicarbonate (374 mg, 1.7 mmol),N,N-dimethylpyridin-4-amine (3.4 mg, 0.029 mmol), andN,N-diethylethanamine (173 mg, 1.7 mmol) in THF (20 mL) was stirredovernight at ambient temperature. The mixture was concentrated in vacuo,and the residue was purified by silica gel chromatography(hexane:EtOAc=100:0-80:20) to give tert-butyl(6′-bromodispiro[cyclobutane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl)carbamate(223 mg).

Preparation Example 24

To a solution of6′-bromodispiro[oxetane-3,3′-chromene-4′,4″-[1,3]thiazol]-2″-amine (214mg, 0.627 mmol) in THF (2.1 mL) were added di-tert-butyl dicarbonate(411 mg, 1.88 mmol) and 4-dimethylaminopyridine (3.8 mg, 0.031 mmol).The mixture was stirred overnight at ambient temperature, anddi-tert-butyl dicarbonate (68.4 mg, 0.314 mmol) was added to thereaction mixture. After stirring for 2 hours at ambient temperature, themixture was partitioned between EtOAc and 10 wt. % aqueous citric acid.The organic layer was washed with brine, dried over MgSO₄ and silica geland filtered. The filtrate was evaporated off, and purification of theresidue with column chromatography on silica gel (Hexane-EtOAc, a lineargradient of EtOAc from 0 to 50%) afforded di-tert-butyl(6′-bromodispiro[oxetane-3,3′-chromene-4′,4″-[1,3]thiazol]-2″-yl)imidodicarbonate(279 mg).

Preparation Example 26

The mixture of 4-bromo-2-iodophenol (3.30 g, 11.04 mmol),1-bromo-5-chloropentan-2-one (75% purity, 3.9 g, 14.66 mmol), and K₂CO₃(2.3 g, 16.64 mmol) in acetone (66 mL) was stirred for 48 hours atambient temperature. The insoluble material was removed by filtrationand washed with EtOAc. The filtrate was evaporated in vacuo. The residuewas purified by silicagel column chromatography (EtOAc-hexane, a lineargradient of EtOAc from 0 to 25%) afforded1-(4-bromo-2-iodophenoxy)-5-chloropentan-2-one (2.12 g).

Preparation Example 27

A solution of potassium tert-butoxide (441 mg, 3.93 mmol) in THF (5 mL)was added to a suspension of6-bromospiro[chromene-2,1′-cyclobutan]-4(3H)-one (500 mg, 1.87 mmol) and1H-benzotriazole-1-methanol (586 mg, 3.93 mmol) in THF (5 mL) over 10minutes in a dry ice-acetone bath under an argon atmosphere. The mixturewas stirred for 0.5 hours in an ice bath, and then diluted with EtOAc(10 mL). After stirring for 0.5 hours, the mixture was filtered off. Thefiltrate was washed with 0.2 M aqueous NaOH (two times), water andbrine, dried over MgSO₄ and silicagel, filtered off. The filtrate wasevaporated to give crude6-bromo-3,3-bis(hydroxymethyl)spiro[chromene-2,1′-cyclobutan]-4(3H)-one(641 mg).

Preparation Example 29

A mixture of di-tert-butyl(6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(300 mg, 0.571 mmol), 3-methoxypyridin-2-amine (354 mg, 2.86 mmol),tris(dibenzylideneacetone)dipalladium(0) (105 mg, 0.114 mmol),(9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine) (198 mg, 0.343mmol), Cs₂CO₃ (558 mg, 1.71 mmol) and dioxane (15 mL) was stirred for 48hours at 100° C. The reaction mixture was cooled down to ambienttemperature, and partitioned with CHCl₃ and water. The organic layer wasdried over Na₂SO₄, and filtered. The filtrate was concentrated atreduced pressure, to give crude di-tert-butyl{6′-[(3-methoxypyridin-2-yl)amino]dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl}imidodicarbonate,which was used for the next reaction without further purification.

Preparation Example 30

To a mixture of(4-amino-6-bromo-4H-spiro[chromene-3,3′-oxetan]-4-yl)methanol (280 mg,0.933 mmol), CH₂Cl₂ (10 mL) and saturated aqueous NaHCO₃ (10 mL) wasadded a mixture of chloroacetyl chloride (0.083 mL, 1.02 mmol) andCH₂Cl₂ (1 mL) at ambient temperature. After stirring for 30 minutes atambient temperature, chloroacetyl chloride (0.016 mL, 0.197 mmol) wasadded to the reaction mixture. The mixture was stirred for 10 minutes atambient temperature, diluted with CH₂Cl₂ and separated. The organiclayer was washed with water, dried over MgSO₄ and filtered.Concentration of the filtrate at reduced pressure gave crudeN-[6-bromo-4-(hydroxymethyl)-4H-spiro[chromene-3,3′-oxetan]-4-yl]-2-chloroacetamide,which was used for the next reaction without further purification.

Preparation Example 31

To a mixture of crudeN-[6-bromo-4-(hydroxymethyl)-4H-spiro[chromene-3,3′-oxetan]-4-yl]-2-chloroacetamide(351 mg, 0.933 mmol) and 2-methylbutan-2-ol (6.3 mL) was added potassiumtert-butoxide (356 mg, 3.17 mmol) at ambient temperature, and themixture was stirred for 1 hour at the same temperature. MeOH (3.2 mL)was added to the reaction mixture, and the mixture was concentrated atreduced pressure. Purification of the residue with column chromatographyon silica gel (Hexane-EtOAc, a linear gradient of EtOAc from 50 to 100%)afforded6′-bromo-5H-dispiro[1,4-oxazinane-3,4′-chromene-3′,3″-oxetan]-5-one (277mg).

Preparation Example 36

To a suspension of methyl(triphenyl)phosphonium bromide (8.13 g, 22.3mmol) in THF (44 mL) was added n-butyllithium (1.65 M in n-hexane, 13.5mL, 22.3 mmol) in a dry ice-acetone bath under argon atmosphere. Themixture was stirred for 60 minutes in an ice bath. To the mixture wasadded a mixture of6-bromo-2,2-dimethyl-4H-spiro[chromene-3,3′-oxetan]-4-one (2.21 g, 7.44mmol) and THF (11 mL) in an ice bath. The mixture was stirred for 1 hourat ambient temperature. The reaction was quenched by adding water in anice-water bath. The mixture was partitioned between EtOAc-hexane (1:2)and water. The organic layer was washed with brine, dried over MgSO₄,filtered, and the filtrate was evaporated. Purification using silica gelcolumn chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0to 20%) afforded6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,3′-oxetane] (2.02g).

Preparation Example 48

To a mixture of 6-bromo-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one (1.00g, 3.92 mmol) and dioxane (10 mL) were added formaldehyde (37 wt. % inwater, 2.95 mL, 39.2 mmol) and Na₂CO₃ (831 mg, 7.84 mmol) at roomtemperature. After stirring overnight at the same temperature, thereaction mixture was filtered. The filtrate was diluted with CHCl₃,washed with 1M aqueous HCl and water, dried over MgSO₄ and filtered. Thefiltrate was concentrated at reduced pressure, and the residue waspurified with column chromatography on silica gel (hexane-EtOAc, alinear gradient of EtOAc from 0 to 50%) to afford6-bromo-3,3-bis(hydroxymethyl)-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one(1.07 g).

Preparation Example 58

To a solution of 1-(4-bromo-2-iodophenoxy)-5-chloropentan-2-one (1.61 g,3.86 mmol) in THF (35 mL) was added vinylmagnesium bromide (1M solutionin THF, 4.3 mL) at −78° C. under an argon atmosphere. After stirring for1 hour at −78° C., the mixture was gradually warmed up to −30° C. over 1hour. The reaction was quenched by adding saturated aqueous NH₄Clsolution and extracted with EtOAc. The combined organic layer werewashed with brine, dried over Na₂SO₄, filtered, and the filtrate wasevaporated. The residue was purified by silicagel column chromatography(EtOAc-hexane, a linear gradient of EtOAc from 0 to 20%) afforded3-[(4-bromo-2-iodophenoxy)methyl]-6-chlorohex-1-en-3-ol (1.35 g).

Preparation Example 59

The mixture of 3-[(4-bromo-2-iodophenoxy)methyl]-6-chlorohex-1-en-3-ol(1.53 g, 3.43 mmol), Pd(OAc)₂ (77 mg, 0.343 mmol), PPh₃ (360 mg, 1.37mmol), and K₂CO₃ (2.84 g, 20.55 mmol) in MeCN (45 mL) was heated at 85°C. for 18 hours under an argon atmosphere. After addition of Pd(OAc)₂(23 mg, 0.102 mmol) and PPh₃ (108 mg, 0.412 mmol), the mixture washeated at 85° C. for 30 hours. The reaction mixture was cooled to roomtemperature, and the insoluble material was removed by filtration andwashed with EtOAc. The filtrate was evaporated in vacuo. The residue waspurified by silicagel column chromatography (EtOAc-hexane, a lineargradient of EtOAc from 0 to 15%) to afford a mixture of6-bromo-4-methylene-4′,5′-dihydro-3′H,4H-spiro[chromene-3,2′-furan] anda reaction intermediate (598 mg). To the mixture dissolved in MeCN (30mL) were added Pd(OAc)₂ (23 mg, 0.102 mmol), PPh₃ (108 mg, 0.412 mmol),and K₂CO₃ (947 mg, 6.85 mmol) and the reaction mixture was heated at 85°C. for 2 hours under an argon atmosphere. The reaction mixture wascooled to room temperature, and the insoluble material was removed byfiltration and washed with AcOEt. The filtrate was evaporated in vacuo.The residue was purified by silicagel column chromatography(EtOAc-hexane, a linear gradient of EtOAc from 0 to 15%) to afford6-bromo-4-methylene-4′,5′-dihydro-3′H,4H-spiro[chromene-3,2′-furan] (460mg).

Preparation Example 60

To a mixture of6′-bromo-5H-dispiro[1,4-oxazinane-3,4′-chromene-3′,3″-oxetan]-5-one (275mg, 0.808 mmol) and dioxane (11 mL) was added2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane 2,4-disulfide (236mg, 0.566 mmol) at ambient temperature. After stirring for 2 hours at80° C., the reaction mixture was cooled down to ambient temperature andconcentrated at reduced pressure. The residue was purified with columnchromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAcfrom 0 to 50%) to afford6′-bromo-5H-dispiro[1,4-oxazinane-3,4′-chromene-3′,3″-oxetane]-5-thione(259 mg).

Preparation Example 62

To a solution of ethyl 1-(hydroxymethyl)cyclobutanecarboxylate (1.0 g,6.3 mmol) and N,N-diethylethanamine (1.5 g, 8.2 mmol) in CH₂Cl₂ (30 ml)was added methanesulfonyl chloride (869 mg, 7.6 mmol). The mixture wasstirred for 6 hours at ambient temperature. After dilution with CHCl₃and H₂O, the organic layer was washed with H₂O, dried over MgSO₄, andconcentrated in vacuo to give ethyl1-{[(methylsulfonyl)oxy]methyl}cyclobutanecarboxylate (1.2 g).

Preparation Example 63

To a mixture of6′-bromo-2H-dispiro[1,3-oxazolidine-4,4′-chromene-3′,3″-oxetan]-2-one(1.98 g, 6.07 mmol), EtOH (9.9 mL) and water (50 mL) was added lithiumhydroxide monohydrate (2.68 g, 60.7 mmol), and the mixture was stirredovernight at 100° C. The reaction mixture was cooled down to ambienttemperature and extracted with CH₂Cl₂. The organic layer was washed withwater, dried over Na₂SO₄ and filtered. After concentration of thefiltrate at reduced pressure, the residue was triturated with hexane,collected by filtration, washed with EtOAc-hexane (1:3) and dried atreduced pressure to afford(4-amino-6-bromo-4H-spiro[chromene-3,3′-oxetan]-4-yl)methanol (1.40 g).

Preparation Example 64

To a solution of di-tert-butyl(6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(300 mg, 0.571 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (174 mg,0.685 mmol), and PdCl₂(dppf) (21 mg, 0.029 mmol) in dioxane (6 mL) wasadded potassium acetate (112 mg, 1.14 mmol). The mixture was stirred for3 hours at 110° C. The resulting precipitate was removed by filtrationand the filtrate was evaporated. Silicagel column chromatography(MeOH—CHCl₃, a linear gradient of MeOH from 3 to 10%) affordeddi-tert-butyl[6′-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(305 mg).

Preparation Example 66

To a mixture of di-tert-butyl(6′-bromodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl)imidodicarbonate(200 mg, 0.393 mmol), bis(dibenzylideneacetone)palladium (0) (22.6 mg,0.039 mmol), and tri-tert-butylphosphonium tetrafluoroborate (11.6 mg,0.039 mmol) was added lithium bis(trimethylsilyl)amide (1 M in toluene,1.96 mL, 1.96 mmol) at ambient temperature. After stirring for 1 hour at100° C., the mixture was cooled down to ambient temperature. To themixture were added 1M aqueous HCl (1.96 mL) and MeOH (1.96 mL) atambient temperature, and the mixture was stirred for 30 minutes at thesame temperature. The mixture was extracted with CHCl₃, and the organiclayer was dried over Na₂SO₄ and filtered. The filtrate was concentratedat reduced pressure and purified with column chromatography on silicagel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%) to affordtert-butyl(6′-aminodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl)carbamate(103 mg).

Preparation Example 67

Under argon atmosphere, to a mixture of di-tert-butyl(6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(1.00 g, 1.81 mmol), bis(dibenzylideneacetone)palladium (0) (104 mg,0.181 mmol) and tri-tert-butylphosphonium tetrafluoroborate (53.5 mg,0.181 mmol) was added lithium bis(trimethylsilyl)amide (1.0 M intoluene, 9.03 mL, 9.03 mmol) at ambient temperature. After stirring for1 hour at 100° C., the reaction mixture was cooled down to ambienttemperature. To the mixture were added 1.0 M hydrochloric acid (9.0 mL)and MeOH (9.0 mL) and the mixture was stirred for 30 minutes at ambienttemperature. After extraction of the mixture with CHCl₃, the organiclayer was dried over Na₂SO₄ and filtered. The filtrate was concentratedat reduced pressure and the residue was purified by columnchromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAcfrom 0 to 90%) to give tert-butyl(6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)carbamate(616 mg).

Preparation Example 69

Under argon atmosphere, a mixture of 4,6-dichloropyrimidine (500 mg,3.36 mmol), 1-(trimethylsilyl)-1-propyne (0.497 mL, 3.36 mmol),tetrabutylammonium fluoride (1 M in THF, 3.36 mL, 3.36 mmol),triethylamine (1.54 mL, 11.1 mmol),tetrakis(triphenylphosphine)palladium(0) (194 mg, 0.168 mmol), copper(I)iodide (192 mg, 1.01 mmol) and toluene (20 mL) was stirred for 9 hoursat 60° C. The mixture was cooled down to ambient temperature, and waterwas added to the mixture. The mixture was extracted with CHCl₃, and theorganic layer was dried over MgSO₄ and filtered. The filtrate wasconcentrated at reduced pressure, and purification of the residue withcolumn chromatography on silica gel (hexane-EtOAc, a linear gradient ofEtOAc from 0 to 20%) afforded 4-chloro-6-(prop-1-yn-1-yl)pyrimidine (207mg).

Preparation Example 70

A mixture of tert-butyl(6′-aminodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl)carbamate(115 mg, 0.333 mmol), 5-fluoropyridine-2-carboxylic acid (62.3 mg, 0.433mmol), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride(83.0 mg, 0.433 mmol), 1-hydroxybenzotriazole (58.5 mg, 0.433 mmol),N,N-diisopropylethylamine (0.074 mL, 0.433 mmol) and CH₂Cl₂ (1.2 mL) wasstirred for 2.5 days at ambient temperature. The reaction mixture waspurified with column chromatography on silica gel (hexane-EtOAc, alinear gradient of EtOAc from 10 to 90%) to afford tert-butyl(6′-{[(5-fluoropyridin-2-yl)carbonyl]amino}dispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl)carbamate(136 mg).

Preparation Example 73

A mixture of tert-butyl(6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)carbamate(878 mg, 2.25 mmol), 5-chloro-2-pyridinecarboxylic acid (476 mg, 2.93mmol), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (562mg, 2.93 mmol), 1-hydroxybenzotriazole (396 mg, 2.93 mmol),N,N-diisopropylethylamine (0.502 mL, 2.93 mmol) and CH₂Cl₂ (8.78 mL) wasstirred for 1.5 hours at ambient temperature. After concentration of thereaction mixture at reduced pressure, the residue was purified withcolumn chromatography on silica gel (hexane-EtOAc, a linear gradient ofEtOAc from 0 to 90%) to give tert-butyl(6′-{[(5-chloropyridin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)carbamate(976 mg).

Preparation Example 76

A mixture of di-tert-butyl(6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(150 mg, 0.286 mmol) and 3-methoxyprop-1-yne (0.072 mL, 0.86 mmol) inEt₃N (1.5 mL) was purged with argon. To the mixture was added Pd(PPh₃)₄(13 mg, 0.011 mmol) and CuBr (4.9 mg, 0.034 mmol), and the mixture wasrefluxed for 3 hours under an argon atmosphere. The mixture waspartitioned between CHCl₃ and brine, and filtered through celite. Theorganic layer of the filtrate was dried over MgSO₄, filtered off, andthe filtrate was evaporated. Silicagel column chromatography(EtOAc-hexane, a linear gradient of EtOAc from 0 to 25%) affordeddi-tert-butyl[6′-(3-methoxyprop-1-yn-1-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(34.7 mg).

Preparation Example 78

The mixture of 6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-one (8.0 g,32 mmol), 2-methylpropane-2-sulfinamide (12 g, 99 mmol), andtitanium(IV) tetraethanolate (22 g, 95 mmol) in THF (160 ml) was stirredfor 48 hours at 80° C. To the mixture was added H₂O (20 ml), filteredthrough Celite and washed by EtOAc (50 ml). The filtrate wasconcentrated in vacuo. The residue was purified by silica gelchromatography (hexane:EtOAc=100:0-0:100) to giveN-(6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-ylidene)-2-methylpropane-2-sulfinamide(6.5 g).

Preparation Example 83

A mixture of di-tert-butyl(6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(300 mg, 0.571 mmol),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyran(360 mg, 1.71 mmol), bis(triphenylphosphine)palladium(II) dichloride(40.1 mg, 0.057 mmol) and Na₂CO₃ (182 mg, 1.71 mmol) in dioxane (3.6 mL)and water (0.9 mL) was stirred for 1 hour at 100° C. The mixture wascooled down to ambient temperature and partitioned between EtOAc andwater. The organic layer was washed with brine, dried over MgSO₄,diluted with hexane, and filtered through silica gel pad (eluted with50% EtOAc in hexane). The filtrate was concentrated at reduced pressureto give crude di-tert-butyl[6′-(3,6-dihydro-2H-pyran-4-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonatewhich was used for the next reaction without further purification.

Preparation Example 85

Under argon atmosphere, a mixture of di-tert-butyl[6′-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(213 mg, 0.372 mmol), 3-bromo-5-(3-methoxyprop-1-yn-1-yl)pyridine (252mg, 1.12 mmol), Na₂CO₃ (158 mg, 1.49 mmol),tetrakis(triphenylphosphine)palladium(0) (21.5 mg, 0.019 mmol), dioxane(3.4 mL) and water (0.85 mL) was stirred for 3 hours at 110° C. Thereaction mixture was cooled down to ambient temperature, and water wasadded to the mixture. The mixture was extracted with MeOH—CHCl₃ (1:9),and the organic layer was dried over Na₂SO₄ prior to filtration. Thefiltrate was concentrated at reduced pressure to give crudedi-tert-butyl{6′-[5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl}imidodicarbonate,which was used for the next reaction without further purification.

Preparation Example 91

A mixture of di-tert-butyl[6′-(5-bromopyridin-3-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(232 mg, 0.385 mmol), ethynyl(trimethyl)silane (0.160 mL, 1.16 mmol),bis(triphenylphosphine)palladium(II) dichloride (13.5 mg, 0.019 mmol),copper(I) iodide (7.3 mg, 0.039 mmol) and triethylamine (3.2 mL) wasstirred overnight at ambient temperature and for 6 days at 50° C.Ethynyl(triisopropyl)silane (0.257 mL, 1.16 mmol) was added to thereaction mixture at ambient temperature, and the mixture was stirredovernight at 85° C. The mixture was cooled down to ambient temperature,diluted with EtOAc and washed with saturated aqueous NH₄Cl. The organiclayer was dried over MgSO₄ and filtered. The filtrate was concentratedat reduced pressure.

To a mixture of the residue and THF (4.6 mL) was addedtetrabutylammonium fluoride (1M in THF, 1.54 mL, 1.54 mmol), and themixture was stirred overnight at ambient temperature. The mixture waspartitioned with EtOAc and saturated aqueous NH₄Cl, and the organiclayer was washed with brine, dried over anhydrous MgSO₄ and filtered.The filtrate was concentrated at reduced pressure, and purification ofthe residue with column chromatography on silica gel (hexane-EtOAc, alinear gradient of EtOAc from 0 to 50%) afforded di-tert-butyl[6′-(5-ethynylpyridin-3-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(59.9 mg).

Preparation Example 92

To a solution of 6-bromo-4H-chromen-4-one (12 g, 53 mmol) in CH₂Cl₂ (24mL) was added TMSOTf (12.5 mL, 69.18 mmol) at ambient temperature. Afterstirring for 1 hour, THF (210 mL) was added to the mixture at ambienttemperature and cooled to −78° C. To the mixture was addedn-propylmagnesium bromide (1.05M solution in THF, 66 mL, 69 mmol). Afterstirring for 1 hour at −78° C., 1M aqueous NH₄Cl was added to themixture. The mixture was warmed to ambient temperature and stirredovernight. The organic and the aqueous layers were separated, and theorganic layer was dried over Na₂SO₄, filtered, and the filtrate wasconcentrated in vacuo. The residue was purified by silicagel columnchromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 10%)to afford 6-bromo-2-propyl-2,3-dihydro-4H-chromen-4-one (11.42 g).

Preparation Example 94

A mixture of 4-(methoxymethyl)-1H-pyrazole (145 mg, 1.29 mmol),6′-bromo-4′H-dispiro[cyclobutane-1,2′-chromene-3′,3″-oxetan]-4′-one (200mg, 0.647 mmol), trans-N,N′-dimethylcyclohexane-1,2-diamine (74 mg, 0.52mmol) and K₂CO₃ (268 mg, 1.94 mmol) in NMP (2 mL) was purged with argon.To the mixture was added CuI (49 mg, 0.26 mmol), and the mixture wassealed and stirred for 1 hour at 150° C. and 0.5 hours at 170° C. undera microwave irradiation. The mixture was partitioned between EtOAc andsaturated aqueous NH₄Cl. The organic layer was washed with water (twotimes) and brine, dried over MgSO₄, filtered, and the filtrate wasevaporated. Silicagel column chromatography (EtOAc-hexane, a lineargradient of EtOAc from 0 to 45%) afforded6′-[4-(methoxymethyl)-1H-pyrazol-1-yl]-4′H-dispiro[cyclobutane-1,2′-chromene-3′,3″-oxetan]-4′-one(97 mg).

Preparation Example 96

To a mixture of6-bromo-3,3-bis(hydroxymethyl)-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one(6.76 g, 21.4 mmol), zinc bis(dimethyldithiocarbamate) (26.2 g, 85.8mmol) and triphenylphosphine (8.44 g, 32.2 mmol) in THF (0.20 L) wasadded diisopropyl azodicarboxylate (1.9 M solution in toluene, 16.9 mL,32.2 mmol) in an ice-water bath. The mixture was stirred overnight atambient temperature. The mixture was diluted with toluene (0.20 L), andthe mixture was filtered off. The filtrate was washed with 1 M aqueousNaOH (three times), water and brine, dried over MgSO₄ and filtered. Thefiltrate was evaporated to give a crude product. The crude product waspurified with column chromatography on silica gel (Hexane-EtOAc, alinear gradient of EtOAc from 0 to 20%) to afford6-bromo-2,2-dimethyl-4H-spiro[chromene-3,3′-oxetan]-4-one (2.22 g).

Preparation Example 106

To a solution of ethyl 1-[(4-bromophenoxy)methyl]cyclobutanecarboxylate(1.1 g, 3.5 mmol) in EtOH (11 ml) was added 1M aqueous NaOH (11 ml, 1mmol). The mixture was stirred for 7 hours at 60° C. The mixture wasconcentrated in vacuo, and to the solution was added 1M aqueous HCl.

The resulting precipitate was collected by filtration, washed with H₂Oand dried in vacuo to give crude1-[(4-bromophenoxy)methyl]cyclobutanecarboxylic acid (0.91 g).

Preparation Example 108

To a mixture of 1-(5-bromo-2-hydroxyphenyl)ethanone (10.0 g, 46.5 mmol)and MeOH (0.20 L) were added 3-methylbutanal (7.54 mL, 69.8 mmol) andpyrrolidine (5.77 mL, 69.8 mmol) at ambient temperature, and the mixturewas stirred for 3 days at the same temperature. After concentration ofthe reaction mixture at reduced pressure, the residue was diluted withEtOAc, acidified to pH 3-4 with 1M aqueous HCl, and extracted withEtOAc. The organic layer was washed with water and brine, dried overMgSO₄, and filtered. After concentration of the filtrate at reducedpressure, purification of the residue with column chromatography onsilica gel (EtOAc-hexane, a linear gradient of EtOAc from 0 to 10%)afforded 6-bromo-2-isobutyl-2,3-dihydro-4H-chromen-4-one (9.02 g).

Preparation Example 111

The mixture of 1-(5-bromo-2-hydroxyphenyl)ethanone (10 g, 46.50 mmol),propionaldehyde (6.7 mL, 93 mmol), pyrrolidine (3.9 mL, 47 mmol), andacetic acid (3.2 mL, 56 mmol) in toluene (20 mL) was heated to 60° C.for 18 hours. After cooling to room temperature, the mixture wasconcentrated in vacuo. The mixture was diluted with diethyl ether and 1Maqueous HCl. The phases were separated. The organic phase was washedwith 1M aqueous NaOH, then brine. The organic phase was dried overNa₂SO₄, filtered, and concentrated in vacuo. Purification with silicagelcolumn chromatography (EtOAc-hexane, a linear gradient of EtOAc from 1to 10%) afforded 6-bromo-2-ethyl-2,3-dihydro-4H-chromen-4-one (3.92 g).

Preparation Example 112

The mixture of ethyl1-{[(methylsulfonyl)oxy]methyl}cyclobutanecarboxylate (1.50 g, 6.3mmol), 4-bromophenol (1.2 g, 7.0 mmol), and caesium carbonate (4.13 g,7.0 mmol) in DMF (15 ml) was stirred for 6 hours at 135° C. Afterdilution with EtOAc and H₂O, the organic layer was washed with H₂O,dried over MgSO₄, and concentrated in vacuo. The residue was purified bysilica gel chromatography (hexane:EtOAc=100:0-70:30) to give ethyl1-[(4-bromophenoxy)methyl]cyclobutanecarboxylate (919 mg).

Preparation Example 114

To sulfuric acid (5 mL) was added1-[(4-bromophenoxy)methyl]cyclobutanecarboxylic acid (1.9 g, 6.6 mmol)at 0° C. in an ice bath. After stirring for 1 hour at room temperature,ice was added to the mixture portionwise. The mixture was diluted withwater, and extracted with EtOAc. The organic layer was washed withbrine, dried over MgSO₄, and concentrated in vacuo. The residue waspurified by silica gel chromatography (hexane:EtOAc=100:0-70:30) to give6-bromo-4H-spiro[chromene-3,1′-cyclobutan]-4-one (649 mg).

Preparation Example 116

To a stirred solution of 3,5-dibromopyridine (251 mg, 1.061 mmol) anddi-tert-butyl(6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-yl)imidodicarbonate(200 mg, 0.354 mmol) in dioxane (1.6 ml) were added4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (93 mg, 0.364mmol), potassium acetate (69 mg, 0.707 mmol) and PdCl₂(PPh₃)₂ (50 mg,0.071 mmol) at room temperature and the mixture was stirred at 100° C.for 8 hours before the starting molecule was completely consumed to givethe corresponding boronate intermediate. To this mixture was addedNa₂CO₃ (150 mg, 1.42 mmol) and H₂O (400 μl) and the mixture was stirredat 100° C. for 6 hours before the boronate intermediate was completelyconsumed. The mixture was cooled to room temperature and evaporated togive a crude, which was purified with column chromatography (EtOAc inhexane=0 to 50%) to give di-tert-butyl[6′-(5-bromopyridin-3-yl)trispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-yl]imidodicarbonate(48 mg).

Preparation Example 117

The mixture of4-bromo-1-[(3,3-dimethoxy-1-vinylcyclobutyl)methoxy]-2-iodobenzene (3.16g, 6.98 mmol) and 1M aqueous HCl (14 mL) in THF (31 mL) was stirred forambient temperature for 1 hour. Then the mixture was stirred for 4 hoursat 50° C. The mixture was cooled to room temperature and added saturatedaqueous NaHCO₃ and extracted with EtOAc. The combined organic extractswere washed with brine, dried over Na₂SO₄, filtered, and evaporatedunder reduced pressure to give3-[(4-bromo-2-iodophenoxy)methyl]-3-vinylcyclobutanone (2.91 g).

Preparation Example 119

To a solution of diisopropylamine (3.2 mL, 22.67 mmol) in THF (45 mL)was added n-BuLi (2.69 M in hexane, 7.7 mL, 20.71 mmol) at −78° C. underargon. The mixture was stirred for 10 minutes at 0° C., then cooled to−78° C. and added a solution of methyl3,3-dimethoxycyclobutanecarboxylate (3.0 g, 17.22 mmol) in THF (10 mL).The mixture was stirred for 30 minutes at −78° C., then added a solutionof acetaldehyde (1.9 mL, 33.86 mmol) in THF (10 mL). The mixture wasstirred for 30 minutes at −78° C., and water was added. The aqueouslayer was extracted with EtOAc. Combined organic layers were washed withbrine, dried over Na₂SO₄, filtered, and evaporated under reducedpressure to give methyl1-(1-hydroxyethyl)-3,3-dimethoxycyclobutanecarboxylate (3.33 g).

Preparation Example 128

Under ice cooling, to a solution of2-(4-amino-6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-yl)-2,2-difluoroethanol(2.24 g, 6.46 mmol) in acetone (45 mL) was added benzoyl isothiocyanate(1.16 g, 7.10 mmol), and the mixture was stirred for 2 hours at roomtemperature and stirred for 13 hours at 40° C. After concentration, theresidue was purified by silica gel chromatography(EtOAc/hexane=1:99-30:70) followed by purification using silica gelchromatography (NH-silicagel, EtOH/CHCl₃=0:100-10:90) to giveN-{[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]carbamothioyl}benzamide(684 mg).

Preparation Example 131

To an ice chilled solution of6-bromo-4-methylene-3′H,4H-spiro[chromene-3,1′-cyclobutan]-3′-one (148mg, 0.53 mmol) in CH₂Cl₂ (4.4 mL) was added DAST (0.20 mL, 1.53 mmol),and the mixture was stirred at room temperature for 4.5 hours. Anotherportion of DAST (0.10 mL, 0.76 mmol) was added to the reaction mixtureand the mixture was stirred at room temperature for 19.5 hours. Themixture was cooled at 0° C. and added to saturated aqueous NaHCO₃ andthe resulting mixture was extracted with CHCl₃. The combined organicextracts were dried over Na₂SO₄, filtered, and concentrated in vacuo.The residue was purified by silica gelchromatography(hexane/EtOAc=100:0-90:10) to give6-bromo-3′,3′-difluoro-4-methylene-4H-spiro[chromene-3,1′-cyclobutane](65 mg).

Preparation Example 132

To a solution ofN-{[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]carbamothioyl}benzamide(340 mg, 0.684 mmol) in MeOH (1.7 mL) was added methylamine (9.8M MeOHsolution, 698 μL, 6.84 mmol). The mixture was stirred for 3 hours atambient temperature. The mixture was concentrated azeotropically withtoluene 3 times to give crude1-[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]thiourea(268 mg).

Preparation Example 133

Under ice cooling, to a solution ofN-[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]-2-methylpropane-2-sulfinamide(4.60 g, 10.5 mmol) in THF-EtOH (50% v/v, 46 mL) was added 4MHCl/dioxane (13.1 mL, 52.5 mmol), and the mixture was stirred for 3hours at room temperature. Under ice cooling, saturated aqueous NaHCO₃,H₂O and brine were added to the mixture, and then the mixture wasextracted with EtOAc. The organic layer was dried over MgSO₄ andconcentrated. The residue was purified by silica gel chromatography(EtOAc/hexane=50:50-100:0) to give2-(4-amino-6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-yl)-2,2-difluoroethanol(2.28 g).

Preparation Example 134

To a solution of methyl 3,3-dimethoxy-1-vinylcyclobutanecarboxylate(2.02 g, 10.09 mmol) in THF (20 mL) was added DIBAL-H (1.04M in toluene,29 mL, 30.16 mmol) at 0° C. under argon, and the mixture was stirred for30 minutes at 0° C. To the mixture was carefully added MeOH (29 mL) andNa₂SO₄.10H₂O (29 g), and the mixture was stirred overnight. The mixturewas filtered and evaporated under reduced pressure. And the residue wasdiluted with hexane/EtOAc=1:1 and filtrated through the pad of silicagel and concentrated in vacuo to give(3,3-dimethoxy-1-vinylcyclobutyl)methanol (1.42 g).

Preparation Example 137

Under ice cooling, to a solution of LiBH₄ (458 mg, 21.0 mmol) in THF (30mL) was added a solution of ethyl{6-bromo-4-[(tert-butylsulfinyl)amino]-4H-spiro[chromene-3,1′-cyclopropan]-4-yl}(difluoro)acetate(5.05 g, 10.5 mmol) in THF (20 mL), and the mixture was stirred for 15minutes at same temperature and stirred for 2 hours at room temperature.After adding H₂O and brine, the mixture was extracted with EtOAc. Theorganic layer was dried over MgSO₄ and concentrated to obtain crudeN-[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]-2-methylpropane-2-sulfinamide.The desired compound (4.6 g) was applied to the next step withoutfurther purification.

Preparation Example 149

To a suspension of activated zinc (3.44 g, 52.5 mmol) in Et₂O-THF (50%v/v, 80 ml) under reflux was slowly added a solution of ethylbromodifluoroacetate (8.00 g, 39.4 mmol) andN-(6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-ylidene)-2-methylpropane-2-sulfinamide(4.68 g, 13.1 mmol) in Et₂O-THF (50% v/v, 80 ml) over 40 minutes, andthe mixture was stirred for 4 hours at the same temperature. Aftercooling, the mixture was filtrated through celite pad and washed withEtOAc. To the filtrate were added saturated aqueous NH₄Cl and EtOAc.After separation, the water layer was extracted with EtOAc. The combinedorganic layer was dried over MgSO₄ and concentrated. The residue waspurified by silica gel chromatography (EtOAc:hexane=20:80-100:0) to giveethyl{6-bromo-4-[(tert-butylsulfinyl)amino]-4H-spiro[chromene-3,1′-cyclopropan]-4-yl}(difluoro)acetate(6.08 g).

Preparation Example 151

To a solution of methyl1-(1-hydroxyethyl)-3,3-dimethoxycyclobutanecarboxylate (3.28 g, 15.03mmol) and pyridine (2.4 mL, 29.83 mmol) in CH₂Cl₂ (65 mL) was addedtrifluoromethanesulfonic anhydride (3.0 mL, 17.86 mmol) at −78° C. Themixture was stirred for 10 minutes and then warmed to 0° C. Afterstirring for 15 minutes at 0° C., DBU (9.0 mL, 60.18 mmol) was added tothe reaction mixture and the resulting mixture was stirred for 1 hour atroom temperature. The mixture was partially evaporated under reducedpressure and filtrated through the pad of silica gel and washed withCH₂Cl₂. The filtrate was washed with saturated aqueous NaHCO₃, driedover Na₂SO₄, filtered, and evaporated. The residue was diluted withhexane/EtOAc=4:1 and added small amount of CHCl₃ and filtrated throughthe pad of silica gel to give methyl3,3-dimethoxy-1-vinylcyclobutanecarboxylate (2.21 g).

Preparation Example 152

Under ice cooling, to a solution ofN-{[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4Hspiro[chromene-3,1′-cyclopropan]-4-yl]carbamothioyl}benzamide (340 mg,0.684 mmol) in CH₂Cl₂ (9 mL) was added1-chloro-N,N,2-trimethylprop-1-en-1-amine (206 mg, 1.54 mmol), and themixture was stirred for 17 hours at room temperature. Ice was added andthe mixture was neutralized by 10% aqueous K₂CO₃ and extracted withCH₂Cl₂. The organic layer was washed with brine and dried over MgSO₄ andconcentrated. The residue was purified by silica gel chromatography(EtOAc/hexane=1:99-40:60) to affordN-(6′-bromo-5″,5″-difluoro-5″,6″-dihydrodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]thiazin]-2″-yl)benzamide(198 mg).

Preparation Example 154

To a solution of(4S)-6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(4.41 g, 12.5 mmol) in THF (44 mL) were added di-tert-butyl dicarbonate(6.54 g, 30.0 mmol) and 4-dimethylaminopyridine (76 mg, 0.62 mmol).After stirring for 16 hours at ambient temperature, the mixture wasconcentrated at reduced pressure. The residue was purified by columnchromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAcfrom 10 to 30%) to afford di-tert-butyl[(4S)-6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(6.95 g).

Preparation Example 155

To a solution of(4′R)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(1.54 g, 4.57 mmol) in tetrahydrofuran (21 ml) were added di-tert-butyldicarbonate (2.49 g, 11.4 mmol) and N,N-dimethylpyridin-4-amine (28 mg,0.23 mmol). The mixture was stirred at room temperature overnight andconcentrated in vacuo. The residue was purified by silica gel columnchromatography (hexane/ethyl acetate=3:1) to afford di-tert-butyl[(4′R)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]imidodicarbonate(1.99 g).

Preparation Example 156

A 3-necked-flask was charged with dimethylsulfoxide (17 ml) andpotassium hydroxide (0.84 g, 15 mmol). Then trimethylsulfoxonium iodide(3.3 g, 15 mmol) was added and the mixture was stirred at roomtemperature for 30 minutes. To this mixture,6-bromo-2,2-dimethyl-3-methylene-2,3-dihydro-4H-chromen-4-one (2.0 g,7.5 mmol) and dimethylsulfoxide (3 ml) were added. The mixture wasstirred at room temperature for 15 hours and then water (30 ml) wasadded. The mixture was extracted with a mixture of hexane (70 ml) andethyl acetate (70 ml). The organic layer was washed with water (50 ml)twice and then with brine (30 ml), dried over anhydrous magnesiumsulfate, and concentrated in vacuo. The residue was purified by silicagel column chromatography (NH-silica gel, hexane/ethylacetate=100:1-20:1) to afford6-bromo-2,2-dimethyl-4H-spiro[chromene-3,1′-cyclopropan]-4-one (1.2 g).

Preparation Example 157

To a mixture of di-tert-butyl[(4S)-6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(3.40 g, 6.14 mmol), bis(dibenzylideneacetone)palladium(0) (353 mg,0.614 mmol), and tri-tert-butylphosphonium tetrafluoroborate (179 mg,0.617 mmol) was added lithium bis(trimethylsilyl)amide (1M solution intoluene, 31 mL, 31 mmol) at ambient temperature under argon atmosphere.After stirring for 1.5 hours at 60° C., the mixture was cooled in anice-water bath and 1M aqueous HCl (31 mL) was added. After stirring for10 minutes at ambient temperature, to the mixture was added CHCl₃, andthe mixture was filtered through a pad of celite. The filtrate wasseparated and the aqueous layer was extracted with CHCl₃. The combinedorganic layer was dried over Na₂SO₄ and filtered. The filtrate wasconcentrated at reduced pressure. To the residue were added MeOH (34 mL)and silica gel (neutral; 17 g) at ambient temperature. After stirringfor 1 hour at 40° C., the mixture was concentrated at reduced pressure.The residue was purified with column chromatography on silica gel(hexane-EtOAc, a linear gradient of EtOAc from 10 to 100%) to affordtert-butyl[(4S)-6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]carbamate(2.38 g).

Preparation Example 158

To a mixture of di-tert-butyl

[(4′R)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]imidodicarbonate(1.98 g, 3.68 mmol), bis(dibenzylideneacetone)palladium(0) (212 mg,0.369 mmol), and tri-tert-butylphosphonium tetrafluoroborate (108 mg,0.371 mmol) was added lithium bis(trimethylsilyl)amide (1M solution intoluene, 18 ml, 18 mmol) at room temperature under argon atmosphere. Themixture was stirred at 60° C. for 2 hours and then quenched withsaturated aqueous ammonium chloride. The mixture was extracted withchloroform. The organic layer was dried over anhydrous magnesiumsulfate, and concentrated in vacuo. The residue was purified by silicagel column chromatography (hexane/ethyl acetate=100:0-50:50-0:100) toafford tert-butyl[(4′R)-6′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]carbamate(1.37 g).

Preparation Example 159

N,N,N′,N′-tetramethylmethanediamine (4.8 g, 47 mmol) was added to asolution of 6-bromo-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one (3.0 g, 12mmol) and acetic acid (0.67 ml, 12 mmol) in tetrahydrofuran (43 ml), andthe mixture was stirred at 70° C. for 24 hours. To the mixture was addedacetic anhydride (4.4 ml, 47 mmol), and the mixture was stirred at 70°C. for 4 hours and concentrated in vacuo. The residue was directlypurified by silica gel column chromatography (hexane/ethyl acetate=20:1)to afford 6-bromo-2,2-dimethyl-3-methylene-2,3-dihydro-4H-chromen-4-one(2.8 g).

Preparation Example 160

To a suspension of methyl(triphenyl)phosphonium bromide (20.8 g, 58.2mmol) in tetrahydrofuran (168 ml) was added n-butyllithium (2.69 Msolution in hexane, 21.6 ml, 58.2 mmol) under dry ice-acetone bathcooling and argon atmosphere. The mixture was stirred for 1 hour at 0°C. To the mixture was added6-bromo-2,2-dimethyl-4H-spiro[chromene-3,1′-cyclopropan]-4-one (8.18 g,29 mmol). The mixture was stirred for 1 hour at 0° C. The reaction wasquenched by adding water and extracted with ethyl acetate. The organiclayer was washed with brine, dried over anhydrous magnesium sulfate, andconcentrated in vacuo. The residue was purified by silicagel columnchromatography (hexane/ethyl acetate=20:1-10:1) to afford6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,1′-cyclopropane](7.69 g).

Preparation Example 161

To a mixture of tert-butyl[(4S)-6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]carbamate(1.00 g, 2.57 mmol) and CH₂Cl₂ (10 mL) were added5-chloro-2-pyridinecarboxylic acid (526 mg, 3.34 mmol),N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (640 mg,3.34 mmol), 1-hydroxybenzotriazole (451 mg, 3.34 mmol) andN,N-diisopropylethylamine (0.571 mL, 3.34 mmol) at ambient temperature.After stirring overnight at the same temperature, the reaction mixturewas purified with column chromatography on silica gel (hexane-EtOAc, alinear gradient of EtOAc from 0 to 90%) and then on NH-silica gel(hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%) to givetert-butyl[(4S)-6′-{[(5-chloropyridin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]carbamate(1.02 g).

Preparation Example 165

To a mixture of tert-butyl[(4′R)-6′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]carbamate(100 mg, 0.268 mmol), 5-methoxypyrazine-2-carboxylic acid (45 mg, 0.30mmol) and 1H-benzotriazol-1-ol (40 mg, 0.29 mmol) in dichloromethane (2mL) was added N-[3-(dimethylamino)propyl]-N′-ethylcarbodiimidehydrochloride (57 mg, 0.30 mmol). The mixture was stirred at roomtemperature for 3 hours and directly purified by silica gel columnchromatography (precolumn: NH-silica gel, main column: neutral silicagel, hexane/ethyl acetate=2:1-1:1-0:1) to afford tert-butyl[(4′R)-6′-{[(5-methoxypyrazin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]carbamate(108 mg).

Preparation Example 167

To an ice-water cooled mixture of tert-butyl[(4′R)-6′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]carbamate(430 mg, 1.15 mmol), 5-(difluoromethyl)pyrazine-2-carboxylic acid (221mg, 1.27 mmol) and 1H-benzotriazol-1-ol (170 mg, 1.26 mmol) inchloroform (8.6 ml) was addedN-[3-(dimethylamino)propyl]-N′-ethylcarbodiimide hydrochloride (244 mg,1.28 mmol). The mixture was stirred at room temperature overnight anddirectly purified by silica gel column chromatography (precolumn: basicsilica gel, main column: neutral silica gel, hexane/ethylacetate=2:1-1:1-0:100) to afford tert-butyl[(4′R)-6′-({[5-(difluoromethyl)pyrazin-2-yl]carbonyl}amino)-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]carbamate(373 mg).

Preparation Example 169

A mixture of 6-bromo-2,3-dihydro-4H-chromen-4-one (0.3 g, 1.3 mmol),paraformaldehyde (0.48 g), L-proline (61 mg, 0.53 mmol), and 0.2 Maqueous sodium hydroxide (6 mL) was stirred at room temperature for 16hours. The mixture was extracted with CHCl₃ and concentrated in vacuo.The residue was purified with silica-gel column chromatography(CHCl₃/MeOH=100:0 to 90:10) to give6-bromo-3,3-bis(hydroxymethyl)-2,3-dihydro-4H-chromen-4-one (0.33 g).

Preparation Example 171

To a solution of 6-bromo-4H-spiro[chromene-3,3′-oxetan]-4-one (143 mg,0.531 mmol) in THF (2 mL) was added Tebbe reagent((C₅H₅)₂TiCH₂ClAl(CH₃)₂,μ-Chloro[di(cyclopenta-2,4-dien-1-yl)]dimethyl(μ-methylene)titaniumaluminum,0.5 M in toluene, 2 mL) under ice-water bath cooling. After the reactionmixture was stirred at the same temperature for 2 hours and then roomtemperature for 4 hours, 1M aqueous NaOH (1 mL) was added. Afterdilution with water and filtration with Celite, the insoluble materialwas washed with CHCl₃. The aqueous phase was extracted with CHCl₃ andcombined organic layer was concentrated in vacuo. The residue waspurified with silica-gel column chromatography to give6-bromo-4-methylene-4H-spiro[chromene-3,3′-oxetane] (0.10 g).

Preparation Example 173

Under nitrogen, MeMgBr solution (2.9 M in 2-methyltetrahydrofuransolution, 3.06 L, 8.88 mol) was diluted by adding into THF (8 L) at 0-5°C. A solution of6-bromo-3,3-bis(hydroxymethyl)-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one(700 g, 2.22 mol) in THF (5 L) was added dropwise via a dropping funnelto the diluted MeMgBr solution maintaining the temperature below 5° C.After the addition was complete, the reaction mixture was allowed towarm to room temperature and stirred for 30 minutes. Then the reactionmixture was heated under reflux overnight. The mixture was cooled toroom temperature followed by cooling with an ice-water bath and 6 Mhydrochloric acid (3.7 L, 22.2 mol) was added dropwise via a droppingfunnel over 30 minutes maintaining the temperature below 10° C. Afterthe addition was complete, the mixture was allowed to warm to roomtemperature and stirred for 20 minutes. The mixture was extracted withtoluene (5 L×2) and the combined extracts were washed with brine, driedover Na₂SO₄ and concentrated under reduced pressure. The residue wastreated with a mixture of hexane/toluene (5:1, 2 L) and the resultingsuspension was stirred for 30 minutes. The precipitate was collected byfiltration, washed with hexane and dried under vacuum to give(6-bromo-2,2-dimethyl-4-methylene-3,4-dihydro-2H-chromene-3,3-diyl)dimethanol(500 g).

Preparation Example 174

Under nitrogen, to a solution of(6-bromo-2,2-dimethyl-4-methylene-3,4-dihydro-2H-chromene-3,3-diyl)dimethanol(800 g, 2.55 mol) and MsCl (877.9 g, 7.66 mol) in THF (4.0 L) was addedEt₃N (851 g, 8.41 mol) over 45 minutes maintaining the temperature below0-10° C. The reaction mixture was allowed to warm to room temperatureand stirred for 1 hour. EtOH (8 L) and NaOH (1021.8 g, 25.55 mol) wereadded and the reaction mixture was heated under reflux for 16 hours.Water (4 L) was added and a clear solution was obtained. Most of thesolvent was removed under reduced pressure and the resulting residue wasextracted with ethyl acetate (1.5 L×2). The combined extracts werewashed with brine, dried over Na₂SO₄ and concentrated under reducedpressure. The residue was dissolved in MeOH (640 mL) at 60° C. and theresulting solution was allowed to cool to room temperature. Theprecipitation formed was filtered-off. The filtrate was concentratedunder reduced pressure and the residue was re-dissolved in MeOH (500 mL)at 60° C. The resulting solution was allowed to cool to room temperatureand then further to 0-5° C. The mixture was stirred at 0-5° C. overnightand the precipitated yellow solid was collected by filtration to give6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,3′-oxetane] (197.2g). The filtrate was concentrated under reduced pressure and the residuewas purified by column chromatography on silica gel (petroleumether/ethyl acetate=50:1) to give another batch of6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,3′-oxetane] (143.6g).

The compounds of Preparation Examples shown in Tables below wereprepared using the respective corresponding starting materials in thesame manner as the methods of Preparation Examples above. The structuresand the preparation methods are shown in [Table. 2] below, and thephysicochemical data for the compounds of Preparation Examples are shownin [Table. 3] below.

TABLE 2 Rf Syn Structure  1 R1

 2 R2

 3 R2

 4 R22

 5 R22

 6 R22

 7 R22

 8 R8 or R22

 9 R22

 10 R22

 11 R22

 12 R22

 13 R22

 14 R22

 15 R22

 16 R22

 17 R22

 18 R22

 19 R22

 20 R22

 21 R22

 22 R22

 23 R23

 24 R24

 25 R24

 26 R26

 27 R27

 28 R29

 29 R29

 30 R30

 31 R31

 32 R36

 33 R36

 34 R36

 35 R36

 36 R36

 37 R36

 38 R36

 39 R36

 40 R36

 41 R36

 42 R36

 43 R36

 44 R36

 45 R36

 46 R36

 47 R36

 48 R48

 49 R48

 50 R48

 51 R48

 52 R48

 53 R48

 54 R48

 55 R48

 56 R48

 57 R48

 58 R58

 59 R59

 60 R60

 61 R62

 62 R62

 63 R63

 64 R64

 65 R64

 66 R66

 67 R66 or R67

 68a R69

 68b R69

 69 R69

 70 R70

 71 R70

 72 R70

 73 R70 or R73

 74 R70

 75 R70

 76 R76

 77 R76

 78 R78

 79 R83

 80 R83

 81 R83

 82 R83

 83 R83

 84 R83

 85 R85

 86 R85

 87 R85

 88 R85

 89 R85

 90 R83

 91 R91

 92 R92

 93 R92

 94 R94

 95 R96

 96 R96

 97 R96

 98 R96

 99 R96

100 R96

101 R96

102 R96

103 R96

104 R96

105 R96

106 R106

107 R106

108 R108

109 R108

110 R108

111 R111

112 R112

113 R112

114 R114

115 R114

116 R116

117 R117

118 R1

119 R119

120 R22

121 R22

122 R22

123 R22

124 R22

125 R22

126 R22

127 R22

128 R128

129 R30

130 R31

131 R131

132 R132

133 R133

134 R134

135 R59

136 R60

137 R137

138 R62

139 R63

140 R70

141 R70

142 R70

143 R70

144 R70

145 R70

146 R70

147 R70

148 R70

149 R149

150 R83

151 R151

152 R152

153 R112

154 R154

155 R155

156 R156

157 R157

158 R158

159 R159

160 R160

161 R161

162 R161

163 R161

164 R165

165 R165

166 R165

167 R167

168 R161

169 R169

170 R96

171 R171

172 R64

173 R173

174 R174

175 R64

176 R85

TABLE 3 Rf Data  1 ESI+: 326, 328  2 ESI+: 519  3 ESI+: 565  4 ESI+:525, 527  5 ESI+: 565, 567  6 ESI+: 539, 541  7 ESI+: 539, 541  8 ESI+:553, 555  9 ESI+: 579, 581  10 ESI+: 553, 555; a compound prepared fromReference Example 9a  11 ESI+: 553, 555; a compound prepared fromReference Example 9b  12 ESI+: 581, 583; a compound prepared fromReference Example 13a  13 ESI+: 581, 583; a compound prepared fromReference Example 13b  14 ESI+: 565, 567; a compound prepared fromReference Example 12a  15 ESI+: 565, 567; a compound prepared fromReference Example 12b  16 ESI+: 567, 569; a compound prepared fromReference Example 11a  17 ESI+: 567, 569; a compound prepared fromReference Example 11b  18 ESI+: 621, 623; a compound prepared fromReference Example 14a  19 ESI+: 539, 541; a compound prepared fromReference Example 1a  20 ESI+: 539, 541; a compound prepared fromReference Example 1b  21 ESI+: 527, 529  22 ESI+: 509, 511  23 ESI+:423, 425  24 ESI+: 541, 543  25 ESI+: 569, 571  26 ESI+: 439, 441 [M +Na]+  27 APCI/ESI+: 327  28 ESI+: 539  29  30 ESI+: 376, 378  31 ESI+:340, 342  32 EI: 306, 308  33 APCI/ESI+: 339  34 APCI/ESI+: 307  35 EI:280, 282  36 EI: 294, 296  37 EI: 264, 266  38 EI: 294, 296  39 ESI+:321, 323  40 ESI+: 309, 311  41 ESI+: 307, 309  42 ESI+: 323, 325  43EI: 362, 364  44 ESI+: 357, 359  45 EI: 268, 270  46 ESI+: 321, 323  47EI: 250, 252  48 ESI+: 315, 317  49 ESI+: 301, 303  50 ESI+: 315, 317 51 ESI+: 341, 343  52 ESI+: 329, 331  53 ESI+: 327, 329  54 ESI+: 365,367 [M + Na]+  55 ESI+: 383, 385  56 ESI+: 377, 379  57 ESI+: 341, 343 58 EL 444, 446  59 ESI+: 281, 283  60 APCI/ESI+: 356, 358  61 ESI+: 209 62 ESI+: 237  63 ESI+: 300, 302  64  65 ESI+: 601  66 ESI+: 346  67ESI+: 390  68a ESI+: 196, 198  68b ESI+: 196, 198  69 ESI+: 153, 155  70ESI+: 469  71 ESI+: 485, 487  72 ESI+: 513  73 ESI+: 529, 531  74 ESI+:547, 549  75 ESI+: 503, 505  76 ESI+: 515  77 ESI+: 561  78 ESI+: 356,358  79 ESI+: 571  80 ESI+: 576  81 ESI+: 525  82 ESI+: 538  83  84APCI/ESI+: 309  85  86  87  88  89 ESI+: 602, 604  90  91 ESI+: 548  92EI: 268, 270  93 EI: 316, 318  94 APCI/ESI+: 341  95 ESI+: 331, 333 [M +Na]+  96 EI: 296, 298  97 ESI+: 297, 299  98 ESI+: 345, 347 [M + Na]+ 99 ESI+: 311, 313 100 ESI+: 331, 333 [M + Na]+ 101 ESI+: 347, 349 [M +Na]+ 102 ESI+: 365, 367 103 ESI+: 359, 361 104 ESI+: 323, 325 105 EI:282, 284 106 EI: 284, 286 107 EI: 288, 290 108 ESI+: 283, 285 109 EI:322, 324 110 ESI+: 281, 283 111 EI: 254, 256 112 EI: 312, 314 113 EI:302, 304 114 EI: 266, 268 115 EI: 270, 272 116 117 EI: 406, 408 118 119ESI+: 241 [M + Na]+ 120 ESI+: 615, 617; a compound prepared fromReference Example 15a 121 ESI+: 615, 617; a compound prepared fromReference Example 15b 122 ESI+: 473, 475 123 ESI+: 579, 581; a compoundprepared from Reference Example 16a 124 ESI+: 559, 561 125 ESI+: 579,581; a compound prepared from Reference Example 16b 126 ESI+: 559, 561127 ESI+: 523, 525 128 ESI+: 497, 499 129 ESI+: 360, 362 130 ESI+: 324,326 131 EI: 300, 302 132 133 134 ESI+: 195 [M + Na]+ 135 EI: 278, 280136 ESI+: 340, 342 137 138 ESI+: 273 [M + Na]+ 139 ESI+: 284, 286 140ESI+: 530, 532 141 ESI+: 574, 576 142 ESI+: 482 143 ESI+: 490 144 ESI+:510, 512 145 ESI+: 499, 501 146 ESI+: 543, 545 147 ESI+: 476 148 ESI+:520 149 ESI+: 480, 482 150 APCI/ESI+: 479 151 ESI+: 223 [M + Na]+ 152ESI+: 479, 481 153 EI: 452, 454 154 ESI+: 553, 555 155 ESI+: 559, 561[M + Na]+ 156 ESI+: 281, 283 157 ESI+: 390 158 ESI+: 374 159 CI+: 267,269 160 ESI+: 279, 281 161 ESI+: 529, 531 162 ESI+: 573, 575 163 ESI+:526 164 ESI+: 513 165 ESI+: 510 166 ESI+: 497 167 ESI+: 530 168 ESI+:526 169 ESI+: 287, 289 170 EI: 268, 270 171 172 ESI+: 613 173 NMR-CDCl₃:1.33 (s, 6H), 2.32 (br s, 2H), 3.90 (d, J = 11.1 Hz, 2H), 4.02 (d, J =11.1 Hz, 2H), 5.51 (s, 1H), 5.86 (s 1H), 6.69 (d, J = 8.7 Hz, 1H),7.31-7.27 (m, 1H), 7.68 (br s, 1H) 174 NMR-CDCl₃: 1.46 (s, 6H), 4.55 (d,J = 6.6 Hz, 2H), 4.71 (d, J = 6.6 Hz, 2H), 5.33 (s, 1H), 5.73 (s, 1H),6.70 (d, J = 8.7 Hz, 1H), 7.28 (dd, J = 2.4, 8.7 Hz, 1H), 7.65 (d, J =2.4 Hz, 1H), 175 ESI+: 557 176 ESI+: 554

Reference Example 1a,b

To an ice chilled solution of6-bromo-4-methylene-4′,5′-dihydro-3′H,4H-spiro[chromene-3,2′-furan] (497mg, 1.77 mmol) in EtOAc (2.5 ml) and MeCN (2.5 ml) was added silvercyanate (397 mg, 2.65 mmol) in an ice bath under an argon atmosphere. Tothe mixture was added iodine (673 mg, 2.65 mmol). After stirring for 30minutes in the ice bath and 30 minutes at ambient temperature, themixture was filtered through celite. The filtrate was washed withsaturated aqueous Na₂S₂O₃. The organic layer was washed with brine,dried over Na₂SO₄, filtered, and the filtrate was evaporated to give anoil. The oil was dissolved in THF (5 mL) and a 2M EtOH solution of NH₃(4.5 ml, 9.0 mmol) was added under ice bath cooling. The mixture wasstirred overnight under ice bath cooling and for 1 hour at 70° C. Afterconcentration in vacuo, the residue was diluted with saturated aqueousNaHCO₃ and extracted with CHCl₃. The organic layer was washed withbrine, dried over Na₂SO₄, filtered, and the filtrate was evaporated off.The residue was purified by silica gelchromatography(CHCl₃/EtOH=100:0-90:10) to give less polar diastereomerof6′-bromo-4,5-dihydro-3H-dispiro[furan-2,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(178 mg) and polar diastereomer of6′-bromo-4,5-dihydro-3H-dispiro[furan-2,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(367 mg).

Reference Example 6

Under an argon atmosphere, to a mixture of6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,3′-oxetane] (504mg, 1.71 mmol), silver cyanate (384 mg, 2.56 mmol) and EtOAc-MeCN (1:1,5.0 mL) was added iodine (649 mg, 2.56 mmol) over 5 minutes in anice-water bath. After stirring for 30 minutes at the same temperature,the mixture was filtered through celite pad. The filtrate was washedwith saturated aqueous Na₂S₂O₃ and brine, dried over MgSO₄ and filtered.After concentration of the filtrate at reduced pressure, the residue wasdissolved in THF (5.0 mL). The solution was added to 2 M EtOH solutionof ammonia (10.7 mL, 21.4 mmol) in an ice-water bath. The mixture wasstirred for 1 hour in the bath and 3 hours at 70° C. After cooling downto ambient temperature, NH-silica gel was added to the reaction mixture,and the mixture was concentrated at reduced pressure. The residue waspurified with column chromatography on silica gel (CHCl₃-EtOH, a lineargradient of EtOH from 0 to 20%) to afford6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(483 mg).

Reference Example 19

A mixture of di-tert-butyl{6′-[5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl}imidodicarbonate(220 mg, 0.372 mmol), silica gel (neutral; 660 mg) and toluene (2.2 mL)was stirred for 3 hours at 100° C. The mixture was cooled down toambient temperature, and concentrated at reduced pressure. Purificationof the residue with column chromatography on silica gel (CHCl₃-MeOH, alinear gradient of MeOH from 0 to 10%) afforded6′-[5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(92.4 mg).

Example 27

A mixture of tert-butyl(6′-{[(5-chloropyridin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)carbamate(217 mg, 0.410 mmol), silica gel (neutral; 651 mg), and toluene (4 mL)was stirred for 80 minutes at 120° C. The reaction mixture was cooleddown to ambient temperature and concentrated at reduced pressure.Purification of the residue with column chromatography on silica gel(eluted with EtOH/CHCl₃=0/100 to 20/80) affordedN-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-chloropyridine-2-carboxamide(144 mg).

Reference Example 31

To a mixture of tert-butyl(6′-{[(5-fluoropyridin-2-yl)carbonyl]amino}dispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl)carbamate(135 mg, 0.288 mmol) and CH₂Cl₂ (5.4 mL) was added trifluoroacetic acid(1.30 mL, 17.0 mmol) at ambient temperature. After stirring for 2 hoursat the same temperature, the mixture was concentrated at reducedpressure. The residue was partitioned between CHCl₃ and saturatedaqueous NaHCO₃. The organic layer was washed with water, dried overNa₂SO₄ and filtered. The filtrate was concentrated at reduced pressure,and the residue was purified with column chromatography on silica gel(CHCl₃-EtOH, a linear gradient of EtOH from 0 to 20%) to affordN-(2″-aminodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl)-5-fluoropyridine-2-carboxamide(43.6 mg).

Reference Example 39

The mixture of di-tert-butyl[6′-(6-methoxypyridin-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(160 mg, 0.289 mmol) and TsOH.H₂O (275 mg, 1.45 mmol) in MeCN (3.2 mL)was stirred for 4 hours at 40° C. After dilution with CHCl₃, the organiclayer was washed with saturated aqueous NaHCO₃ and brine, dried overNa₂SO₄, and concentrated in vacuo. The residue was purified by silicagel chromatography (CHCl₃:MeOH=99:1-95:5) and then the residue waswashed with EtOAc to afford6′-(6-methoxypyridin-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(40 mg).

Reference Example 40

To a solution of 2-fluoro-6-iodobenzonitrile (1560 mg, 6.32 mmol) in THF(16 mL) at −78° C. was added n-butyllithium (2.64M solution in n-hexane,2.39 ml, 6.32 mmol) dropwise. The mixture was stirred for 0.5 hour at−78° C. and to the solution was added a solution ofN-(6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-ylidene)-2-methylpropane-2-sulfinamide(1500 mg, 4.21 mmol) in THF (5 mL). The mixture was stirred for 1 hourat −78° C. and overnight at room temperature. To the mixture was addedsaturated aqueous NH₄Cl. The mixture was extracted with EtOAc and theorganic layer was concentrated in vacuo. The residue was purified bysilica gel chromatography(hexane/AcOEt=100:0-0:100, thenCHCl₃:MeOH=85:15) to give6′-bromo-4″-fluorodispiro[cyclopropane-1,3′-chromene-4′,1″-isoindol]-3″-amine(469 mg).

Reference Example 41

6-Bromo-4-methylene-4H-spiro[chromene-3,3′-oxetane] (1.50 g, 5.62 mmol)was added to a mixture of silver thiocyanate (3.73 g, 22.5 mmol), iodine(2.85 g, 11.2 mmol) and toluene (15 mL) in an ice-water bath. Afterstirring overnight at ambient temperature, the mixture was filteredthrough celite pad (washed with EtOAc). The filtrate was washed withsaturated aqueous Na₂S₂O₃ and brine, dried over MgSO₄ and filtered. Thefiltrate was concentrated at reduced pressure. A mixture of the residueand THF (15 mL) was added to ammonia (2.0 M in EtOH, 30 mL) in anice-water bath. After stirring for 1 hour at the same temperature, themixture was stirred for 2.5 days at ambient temperature. The mixture waspartitioned with MeOH—CHCl₃ (1:9) and water. The organic layer was driedover Na₂SO₄ and filtered. The filtrate was concentrated at reducedpressure, and purification of the residue with column chromatography onsilica gel (CHCl₃-EtOH, a linear gradient of EtOH from 0 to 5%) afforded6′-bromodispiro[oxetane-3,3′-chromene-4′,4″-[1,3]thiazol]-2″-amine (1.26g).

Reference Example 43

The mixture of di-tert-butyl(6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(150 mg, 0.286 mmol), 3-chloropyridin-2-amine (184 mg, 1.43 mmol),(1E,4E)-1,5-diphenylpenta-1,4-dien-3-one-palladium (3:2) (52 mg, 0.057mmol), di-tert-butyl(2′,4′,6′-triisopropylbiphenyl-2-yl)phosphine (97mg, 0.23 mmol), and Cs₂CO₃(279 mg, 0.857 mmol) in 1,4-dioxane (7.5 ml)was stirred for 48 hours at 100° C. The reaction mixture was cooled downto ambient temperature, and partitioned with CHCl₃ and water. Theorganic layer was dried over Na₂SO₄, and filtered. The residue wasdissolved in toluene, and was added silica gel (neutral), and stirred at130° C. for 3 hours. The mixture was concentrated under reducedpressure, and purified by silica gel columnchromatography(CHCl₃/MeOH=100:0-85:15) to give an amorphous, which waswashed with CHCl₃/hexane to give6′-(3-chloropyridin-2-yl)aminodispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetane]-2-amine(28 mg).

Reference Example 46

To a stirred mixture of palladium (II) acetate (7.9 mg, 0.035 mmol) andbiphenyl-2-yl(di-tert-butyl)phosphine (5.3 mg, 0.018 mmol) in THF (5 mL)under argon atmosphere was added di-tert-butyl(6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-yl)imidodicarbonate(100 mg, 0.177 mmol) followed by isobutylzinc bromide (0.5 M THFsolution, 1.1 mL). The reaction mixture was stirred at room temperaturefor 17 hours, and then the reaction was quenched with H₂O and brine. Theresulting mixture was extracted with chloroform 3 times. The combinedorganic layer was dried over magnesium sulfate and concentrated underreduced pressure. The crude product was dissolved in toluene (6 mL). Tothe solution was added silica gel (neutral; 600 mg), and the resultingsuspension was stirred at 100° C. for 1 hour. After cooling andconcentration, the residue was purified by silica gel columnchromatography (EtOH/CHCl₃=0:100-20:80) followed by columnchromatography (NH-silica gel, EtOAc/hexane=20:80-100:0) to afford6′-isobutyltrispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-amine(20 mg).

Reference Example 48

To a stirred solution of di-tert-butyl[6′-(5-bromopyridin-3-yl)trispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-yl]imidodicarbonate(48 mg, 0.075 mmol) in Et₃N (0.67 ml) were added ethynylcyclopropane(0.063 mL, 0.75 mmol), PdCl₂(PPh₃)₂ (10 mg, 0.015 mmol) and CuI (5.7 mg,0.030 mmol) at room temperature and the mixture was sealed andirradiated with microwave at 150° C. for 1 hour. To this mixture wasadded activated carbon (ca. 100 mg) and the mixture was stirred for 1hour to remove palladium residues. The mixture was passed through a padof Celite. The filtrate was evaporated to give a residue. To thesolution of the residue in toluene (3 ml) was added silica gel (500 mg)and the mixture was refluxed for 2 hours. The mixture was passed througha pad of Celite and the filtrate was evaporated to give a crude product,which was purified with column chromatography (MeOH in CHCl₃=0 to 10%)to give 6′-[5-(cyclopropylethynyl)pyridin-3-yl]trispiro[cyclobutane-1,2′chromene-4′,4″-[1,3]oxazole-3′,3′″-o xetan]-2″-amine (5 mg).

Reference Example 49

A suspension of6′-(2-methylimidazo[1,2-a]pyridin-6-yl)trispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-amine(66 mg, 0.16 mmol), acetic acid (27 μl, 0.06 mmol) and PtO₂ (13 mg) inEtOH (3.3 ml) was stirred at room temperature under the hydrogenatmosphere at 3 atm for 8 hours. To the mixture was added acetic acid(63 μl), and the mixture was stirred at room temperature under hydrogenatmosphere at 3 atm for 38 hours. The mixture was filtrated throughcelite pad. And then the filtrate was concentrated under reducedpressure. The obtained residue was purified by silicagel columnchromatography (28% NH₄OH/EtOH/CHCl₃=2:20:80) followed by trituration iniPr₂O and filtration to give6′-(2-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-6-yl)trispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-amine(56 mg).

Reference Example 51a,b

2′,2′-Dimethyl-6′-(pyrimidin-5-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(489 mg, 1.39 mmol) was subjected to chromatography using supercriticalCO₂/[MeOH with 0.1% diethylamine] (70:30) on Chiralcel OD-H column(Daicel, 10×250 mm) eluting at a flow rate 10 mL/minute (40° C. columntemperature). The first peak (retention time=3.44 minutes) provided anenantiomer of2′,2′-dimethyl-6′-(pyrimidin-5-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(220 mg), and the second peak (retention time=6.92 minutes) provided theother enantiomer of2′,2′-dimethyl-6′-(pyrimidin-5-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(215 mg).

Reference Example 52a,b

6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(330 mg, 0.96 mmol) was subjected to chromatography using supercriticalCO₂/[EtOH with 0.1% diethylamine] (65/35) on Chiralcel OZ-H column(10×250 mm) eluting at a flow rate 10 mL/minute (40° C. columntemperature). The first peak (retention time=3.62 minutes) wasconcentrated in vacuo, and dissolved in EtOAc (3 mL) and then hexane (20mL) was added. The resulting precipitate was collected by filtration,washed with hexane and dried in vacuo to give an enantiomer of6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(108 mg). The second peak (retention time=6.27 minutes) was concentratedin vacuo, and dissolved in EtOAc (3 mL) and hexane (20 mL) was added.The resulting precipitate was collected by filtration, washed withhexane and dried in vacuo to give the other enantiomer of6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(80 mg).

Reference Example 53a,b

6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(400 mg, 1.39 mmol) was subjected to chromatography using supercriticalCO₂/[MeOH with 0.1% diethylamine) (70:30) on Chiralcel AD-H column(10×250 mm) eluting at a flow rate 10 mL/minute (40° C. columntemperature). The first peak (retention time=5.81 minutes) provided anenantiomer of6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(160 mg), and the second peak (retention time=9.25 minutes) provided theother enantiomer of6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(170 mg)

Reference Example 54

A mixture of di-tert-butyl(6′-bromo-2′,2′-dimethyldispiro[oxetane-3,3′-chromene-4′,4″-[1,3]thiazol]-2″-yl)imidodicarbonate(268 mg, 0.471 mmol), pyrimidin-5-ylboronic acid (175 mg, 1.41 mmol),bis(triphenylphosphine)palladium(II) dichloride (33.0 mg, 0.047 mmol)and Na₂CO₃ (150 mg, 1.41 mmol) in dioxane-water (4:1, 5.4 mL) wasstirred for 1.5 hours at 100° C.Tetrakis(triphenylphosphine)palladium(0) (272 mg, 0.235 mmol) was addedto the mixture and the mixture was stirred for 1.5 hours at 100° C. Tothe mixture was added charcoal and the mixture was stirred for 30minutes at 50° C. The mixture was filtered through celite pad (elutedwith EtOAc) and the filtrate was washed with water and brine. Theorganic layer was dried over MgSO₄ and filtered, and the filtrate wasconcentrated at reduced pressure. To the residue were added toluene (2.7mL) and silica gel (neutral; 804 mg), and the mixture was stirred for 1hour at 110° C. After concentration of the reaction mixture at reducedpressure, the residue was purified with column chromatography on silicagel (CHCl₃-EtOH, a linear gradient of EtOH from 0 to 20%), thenre-purified with column chromatography on amino silica gel(Hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%). The purifiedproduct was dissolved in dioxane (5.0 mL) and HCl (4 M in dioxane, 0.049mL, 0.198 mmol) was added. The mixture was stirred for 2 hours atambient temperature. After concentration at reduced pressure, theresidue was triturated with IPE, collected by filtration and washed withIPE. The solid was dried under reduced pressure at 30° C. to afford2′,2′-dimethyl-6′-(pyrimidin-5-yl)dispiro[oxetane-3,3′-chromene-4′,4″-[1,3]thiazol]-2″-aminehydrochloride (62.6 mg).

Reference Example 55

Under argon atmosphere, to a mixture of di-tert-butyl(6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(300 mg, 0.571 mmol), dioxane (3.0 mL) and water (1.5 mL) were added1H-indazol-4-ylboronic acid (185 mg, 1.14 mmol), K₂CO₃ (237 mg, 1.71mmol) and bis(triphenylphosphine)palladium(II) dichloride (40 mg, 0.057mmol), and the mixture was stirred for 3 hours at 100° C. The reactionmixture was cooled down to ambient temperature, partitioned between H₂Oand 10% MeOH in CHCl₃. The organic layer was dried over Na₂SO₄ andfiltered, and the filtrate was concentrated at reduced pressure. Theresidue was dissolved with dioxane (3.0 mL), and silica gel (neutral;900 mg) was added to the mixture. After stirring for 2.5 hours at 110°C., the reaction mixture was cooled down to ambient temperature, andconcentrated at reduced pressure. Purification of the residue withcolumn chromatography on silica gel (CHCl₃-EtOH, a linear gradient ofEtOH from 0 to 20%) afforded6′-(1H-indazol-4-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(142 mg).

Reference Example 61

The mixture of di-tert-butyl[6′-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]imidodicarbonate(140 mg, 0.25 mmol), 5-bromo-1H-pyrrolo[2,3-b]pyridine (87 mg, 0.44mmol), Pd(PPh₃)₄ (58 mg, 0.050 mmol), and Na₂CO₃ (80 mg, 0.76 mmol) in1,4-dioxane-H₂O (2.8 ml, 4:1) was stirred for 7 hours at 100° C. Afterdilution with EtOAc and H₂O, the organic layer was washed with H₂O,dried over MgSO₄, and concentrated in vacuo. The residue was purified bysilica gel chromatography (CHCl₃:MeOH=100:0-85:15) to give white solid.The solid was washed by diisopropyl ether to give6′-(1H-pyrrolo[2,3-b]pyridin-5-yl)dispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(42 mg).

Reference Example 65

A mixture of di-tert-butyl[2′,2′-dimethyl-6′-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]imidodicarbonate(282 mg, 0.470 mmol), 2-bromo-4-methoxypyridine (221 mg, 1.17 mmol),tetrakis(triphenylphosphine)palladium(0) (271 mg, 0.235 mmol) and Na₂CO₃(149 mg, 1.41 mmol) in dioxane-water (4:1, 5.6 mL) was stirred for 8hours at 100° C. The mixture was diluted with MeOH—CHCl₃ (1:9) andwashed with water, then the organic layer was dried over MgSO₄ andfiltered. The filtrate was concentrated at reduced pressure. To theresidue were added toluene (2.8 mL) and silica gel (neutral; 846 mg),and the mixture was stirred for 3 hours at 120° C. After concentrationof the reaction mixture at reduced pressure, the residue was purifiedwith column chromatography on silica gel (CHCl₃-EtOH, a linear gradientof EtOH from 0 to 20%) afforded6′-(4-methoxypyridin-2-yl)-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(146 mg).

Reference Example 75

A mixture of di-tert-butyl(6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-yl)imidodicarbonate(300 mg, 0.531 mmol), 5-methylpyridine-3-boronic acid (145 mg, 1.06mmol), bis(triphenylphosphine)palladium(II) dichloride (37 mg, 0.053mmol) and Na₂CO₃ (169 mg, 1.59 mmol) in dioxane-H₂O (4:1, 6.0 mL) wasstirred for 1 hour at 100° C. To the mixture was added charcoal and themixture was stirred for 10 minutes at 50° C. The mixture was filteredthrough celite pad (eluted with EtOAc) and the filtrate was washed withH₂O and brine. The organic layer was dried over MgSO₄ and filtered, andthe filtrate was concentrated under reduced pressure. To the residuewere added toluene (3.0 mL) and silica gel (neutral; 900 mg), and themixture was stirred for 3 hours at 120° C. After concentration of thereaction mixture at reduced pressure, the residue was purified withcolumn chromatography on silica gel (CHCl₃-EtOH, a linear gradient ofEtOH from 0 to 20%) to afford6′-(5-methylpyridin-3-yl)trispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-amine(151 mg).

Reference Example 108

The mixture of6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine (650 mg,2.00 mmol), [5-(prop-1-yn-1-yl)pyridin-3-yl]boronic acid (644 mg, 4.00mmol), (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one-palladium (3:2) (92 mg,0.10 mmol), dicyclohexyl(2′,6′-dimethoxybiphenyl-2-yl)phosphine (164 mg,0.400 mmol), and K₃PO₄ (1.70 g, 8.00 mmol) in DMF (13 mL) was stirredfor 16 hours at 110° C. under argon atmosphere. The precipitate formedwas removed by filtration with celite and washed with CHCl₃. Thefiltrate was concentrated in vacuo. The residue was purified by silicagel chromatography (CHCl₃:MeOH=99:1-95:5) to give6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(310 mg).

Reference Example 114

A mixture of6′-bromo-5H-dispiro[1,4-oxazinane-3,4′-chromene-3′,3″-oxetane]-5-thione(200 mg, 0.561 mmol) and ammonia (2 M in EtOH, 6.0 mL) was stirred for 1week at ambient temperature. The reaction mixture was concentrated atreduced pressure, and the residue was purified with columnchromatography on amino silica gel (CHCl₃-EtOAc, a linear gradient ofEtOAc from 0 to 50% then CHCl₃-EtOH, a linear gradient of EtOH from 0 to20%) to afford6′-bromo-6H-dispiro[1,4-oxazine-3,4′-chromene-3′,3″-oxetan]-5-amine (123mg).

Reference Example 115

A mixture of di-tert-butyl(6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-yl)imidodicarbonate(300 mg, 0.531 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (150 mg,0.584 mmol), potassium acetate (91.1 mg, 0.928 mmol) andbis(triphenylphosphine)palladium(II) dichloride (37 mg, 0.053 mmol) indioxane (2.4 mL) was stirred for 7 hours at 100° C. To the mixture wasadded 3-bromo-2-cyanopyridine (243 mg, 1.33 mmol), Na₂CO₃ (225 mg, 2.12mmol) and water (0.60 mL), and the mixture was stirred for 2 hours at100° C. The mixture was treated with charcoal, and filtered off. Thefiltrate was partitioned between EtOAc and water. The organic layer waswashed with brine, dried over MgSO₄, filtered, and the filtrate wasevaporated to give a pale brown oil. The oil was dissolved in toluene(3.0 mL) and to the mixture was added silica gel (neutral; 900 mg). Themixture was refluxed for 1 hour. The solvent was evaporated off. Silicagel column chromatography (CHCl₃-EtOH, a linear gradient of EtOH from 0to 10%) afforded a solid. The solid was triturated in Et₂O, collected byfiltration, washed with Et₂O and dried at reduced pressure at 70° C. togive3-(2″-aminotrispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-6′-yl)pyridine-2-carbonitrile(141 mg).

Reference Example 116

A mixture of 3-bromo-2-fluoro-5-methylpyridine (202 mg, 1.06 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (300 mg, 1.17mmol), potassium acetate (208 mg, 2.12 mmol) and PdCl₂(PPh₃)₂ (37 mg,0.053 mmol) in dioxane (4 mL) was stirred for 3 hours at 100° C. To themixture was added di-tert-butyl(6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-yl)imidodicarbonate(300 mg, 0.531 mmol), sodium carbonate (225 mg, 2.12 mmol) and H₂O (1mL), and the mixture was stirred for 1.5 hours at 100° C. The mixturewas treated with charcoal, and filtered off. The filtrate waspartitioned between EtOAc and water. The organic layer was washed withbrine, dried over MgSO₄, filtered, and the filtrate was evaporated togive a pale brown oil. The oil was dissolved in toluene (5 mL) and tothe mixture was added silicagel (neutral; 600 mg). The mixture wasrefluxed for 1 hour. The solvent was evaporated off. Silicagel columnchromatography (CHCl₃-EtOH, linear gradient of EtOH from 0 to 20%)afforded a solid. The solid was triturated in Et₂O and collected byfiltration, washed with Et₂O and dried in vacuo at 70° C. to give6′-(2-fluoro-5-methylpyridin-3-yl)trispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-amine(130 mg).

Reference Example 136

A mixture of di-tert-butyl(6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-yl)imidodicarbonate(14.1 mg), 2-fluoro-5-methoxyphenylboronic acid (12.7 mg), PdCl₂(PPh₃)₂(5.3 mg) and 1M aqueous Na₂CO₃ (0.1 mL) in dioxane (0.4 mL) was stirredfor 2 hours at 100° C. The mixture was filtered by using Chem Elutcartridges (Agilent Technologies) and washed with CHCl₃. The filtratewas evaporated to give a brown oil. The oil was dissolved in toluene(0.5 mL) and to the mixture was added silicagel (neutral; 50 mg). Themixture was stirred for 1 hour at 100° C. The mixture was filtered andwashed with CHCl₃. The filtrate was evaporated. The residue was purifiedwith HPLC (Column: Waters SunFire™ Prep C₁₈ OBD™ 5 micrometer, 19×100mm; MeOH/0.1% aqueous HCOOH 11/89 to 95/5(v/v)) and afforded6′-(2-fluoro-5-methoxyphenyl)trispiro[cyclobutane-1,2′-chromene-4′,4″-[1,3]oxazole-3′,3′″-oxetan]-2″-amine(1.5 mg).

Reference Example 146

A mixture of di-tert-butyl(6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(13.8 mg), 3-chloro-5-fluorophenylboronic acid (8.7 mg), Pd(PPh₃)₄ (2.9mg) and 1M aqueous Na₂CO₃ (0.063 mL) in dioxane (0.25 mL) was stirredfor 12 hours at 100° C. The mixture was filtered by using Chem Elutcartridges and washed with CHCl₃. The filtrate was evaporated. Theresidue was purified with HPLC (Column: Waters SunFire™ Prep C₁₈ OBDmicrometer, 19×100 mm; MeOH/0.1% aqueous HCOOH 11/89 to 95/5(v/v)) andafforded6′-(3-chloro-5-fluorophenyl)-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(4.6 mg).

Reference Example 174

A mixture of di-tert-butyl(6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)imidodicarbonate(13.8 mg), indole-5-boronic acid (8.1 mg), PdCl₂(dppf) (2.0 mg) and 1Maqueous K₂CO₃ (0.063 mL) in dioxane (0.25 mL) was stirred for 12 hoursat 100° C. The mixture was filtered by using Chem Elut cartridges andwashed with CHCl₃. The filtrate was evaporated. The residue was purifiedwith HPLC (Column: Waters SunFire™ Prep C₁₈ OBD™ 5 micrometer, 19×100mm; MeOH/0.1% aqueous HCOOH 11/89 to 95/5(v/v)) and afforded6′-(1H-indol-5-yl)-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(4.5 mg).

Reference Example 198

To a solution ofN-[5″,5″-difluoro-6′-(pyrimidin-5-yl)-5″,6″-dihydrodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]thiazin]-2″-yl]benzamide(76 mg, 0.16 mmol) in ethanol (4 mL) were added N-hydroxymethanaminehydrochloride (133 mg, 1.59 mmol) and pyridine (126 mg, 1.59 mmol), andthe mixture was stirred for 27 hours at 70° C. After cooling, H₂O andbrine were added and the mixture was extracted with chloroform. Theorganic layer was dried over MgSO₄ and concentrated. The residue waspurified by silica gel chromatography (EtOAc/hexane=50:50-100:0 and thenEtOH/CHCl₃=5:95) and a resultant solid was washed with iPr₂O to give5″,5″-difluoro-6′-(pyrimidin-5-yl)-5″,6″-dihydrodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]thiazin]-2″-amine(36 mg).

Reference Example 214

A suspension of6′-bromo-5″H-dispiro[cyclopropane-1,3′-chromene-4′,3″-[1,4]oxazinane]-5″-thione(391 mg, 1.15 mmol) in 2M solution of ammonia in ethanol (60 mL, 120mmol) was treated with tert-butylhydroperoxide (5.0-6.0 M solution indecane, 4.60 mL). The mixture was stirred at room temperature for 1hour, followed by the addition of methanol (10 mL). After stirring atroom temperature for 5 hours, insoluble material was filtered off. Thefiltrate was poured into saturated aqueous sodium hydrogen carbonate andextracted with chloroform. The organic layer was washed three times withsaturated aqueous sodium hydrogen carbonate, dried over sodium sulfate,and evaporated. The residue was purified by silica gel chromatography(NH-silica gel, hexane:EtOAc=100:0-0:100) to give6′-bromo-6″H-dispiro[cyclopropane-1,3′-chromene-4′,3″-[1,4]oxazin]-5″-amine(247 mg).

Reference Example 215

In the same manner as in the method of Preparation Example 83 andReference Example 31,6′-(pyrimidin-5-yl)-6″H-dispiro[cyclopropane-1,3′-chromene-4′,3″-[1,4]oxazin]-5″-aminewas prepared with using di-tert-butyl(6′-bromo-6″H-dispiro[cyclopropane-1,3′-chromene-4′,3″-[1,4]oxazin]-5″-yl)imidodicarbonateas a starting material.

Reference Example 216

To a solution of1-[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]thiourea(268 mg, 0.682 mmol) in MeOH (2.7 mL) were added methyliodide (967 mg,6.82 mmol) and 1M aqueous NaOH (0.68 mL, 0.68 mmol). The mixture wasstirred for 3 hours at 60° C. After cooling, to the mixture were addedH₂O and brine. The mixture was extracted with CH₂Cl₂. The extract wasdried over MgSO₄ and concentrated.

Purification using silicagel column chromatography (EtOAc-hexane, alinear gradient of EtOAc from 1 to 100%) afforded a solid (177 mg). To asolution of the solid in MeOH (2.7 mL) was added 1M aqueous NaOH (0.68mL, 0.68 mmol). The mixture was stirred for 5 hours at 60° C. Aftercooling, to the mixture were added H₂O and brine. The mixture wasextracted with EtOAc. The extract was dried over MgSO₄ and concentrated.Purification using silicagel column chromatography (EtOAc-hexane, alinear gradient of EtOAc from 1 to 100%) afforded6′-bromo-5″,5″-difluoro-5″,6″-dihydrodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazin]-2″-amine(145 mg).

Reference Example 217

To a solution of6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,1′-cyclopropane](6.72 g, 24 mmol) in ethyl acetate (67.2 ml) and acetonitrile (67.2 ml)was added silver cyanate (5.4 g, 36 mmol) under cooling with an ice bathunder an argon atmosphere. To the mixture was added iodine (9.17 g, 36mmol). The mixture was stirred at 0° C. for 2 hours and filtered. Thecake was washed with ethyl acetate and the filtrate was partitionedbetween ethyl acetate and saturated aqueous sodium thiosulfate. Theorganic layer was washed with brine, dried over anhydrous magnesiumsulfate, and concentrated in vacuo.

The residue was dissolved in tetrahydrofuran (69 mL) and the solutionwas added to 2 M ethanolic ammonia (151 mL, 302 mmol) under cooling withan ice bath. The mixture was stirred at room temperature overnight, andconcentrated in vacuo.

The residue was dissolved in MeOH (30 ml) and saturated aqueous sodiumbicarbonate was added. The mixture was stirred at room temperature for 1hour and the resulting precipitate was collected and dried in vacuo. Theresidue was triturated with a mixture of ethyl acetate/diisopropyl etherand filtered to afford6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(8.12 g).

Example 218

A mixture of tert-butyl[(4S)-6′-{[(5-chloropyridin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl]carbamate(1.02 g, 1.93 mmol), silica gel (neutral; 3.06 g) and toluene (20.4 mL)was stirred for 3 hours at 120° C. The reaction mixture was cooled downto ambient temperature and concentrated at reduced pressure. The residuewas purified with column chromatography on silica gel (CHCl₃-EtOH, alinear gradient of EtOH from 0 to 15%) and then with NH-silica gel(Hexane-EtOAc, a linear gradient of EtOAc from 50 to 100%). The purifiedproduct was recrystallized from EtOH/H₂O (1:1), and the solid wascollected by filtration and dried at reduced pressure to give a hydrateofN-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide(547 mg) as a crystal.

Example 223

To a solution of tert-butyl[(4′R)-6′-{[(5-methoxypyrazin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]carbamate(392 mg, 0.769 mmol) in chloroform (3 ml) was added trifluoroacetic acid(1.8 ml), and the mixture was stirred at room temperature for 2 hours.The mixture was concentrated in vacuo, and the residue was purified bysilica gel column chromatography (precolumn: NH-silica gel, main column:neutral silica gel, chloroform/methanol=10:0-10:1). To the purifiedproduct was added a mixture of hexane/ethyl acetate (4:1) and themixture was stirred at room temperature over night. The precipitate wascollected, washed with mixture of hexane/ethyl acetate (4:1), and driedin vacuo to affordN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide(246 mg).

Example 224

To a solution of tert-butyl[(4′R)-6′-({[5-(difluoromethyl)pyrazin-2-yl]carbonyl}amino)-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]carbamate(373 mg, 0.704 mmol) in chloroform (6 ml) was added trifluoroacetic acid(2 ml). The mixture was stirred at room temperature for 2 hours andconcentrated in vacuo. The residue was purified by silica gel columnchromatography (precolumn: NH-silica gel, main column: neutral silicagel, chloroform/methanol=100:0-10:1). To the purified product was addeda mixture of hexane and ethyl acetate (4:1), and the mixture was stirredat room temperature over night. The resulting precipitate was collected,washed with a mixture of hexane and ethyl acetate (4:1), and dried invacuo to affordN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide(253 mg).

Reference Example 225a,b

To a solution of6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(3.7 g, 11 mmol) in methanol (50 ml) was added (+)-dibenzoyl-D-tartaricacid monohydrate (4.1 g, 11 mmol). The mixture was stirred at roomtemperature for 5 minutes and concentrated in vacuo. To the residue wasadded dioxane (25 ml). The mixture was heated under reflux for 5minutes, cooled to room temperature and stirred at room temperatureovernight. The resulting precipitate was collected, washed with dioxane,and dried in vacuo. The resulting powder was dissolved in saturatedaqueous sodium hydrogen carbonate and chloroform. The mixture wasextracted with chloroform, and the organic layer was dried overanhydrous magnesium sulfate, and concentrated in vacuo to afford(4′R)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(1.5 g). The mother liquor was concentrated in vacuo and purified byNH-silica gel column chromatography (CHCl₃/MeOH=20:1). The purifiedmaterial was treated with (−)-dibenzoyl-L-tartaric acid monohydrate (2.0g, 5.4 mmol) in the same manner as described above which lead toisolation of(4′S)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-amine(1.2 g).

Reference Example 226

A mixture of racemic6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(155 g, 0.44 mol) and L-camphorsulfonic acid (102 g, 0.44 mol) inethanol (2.7 L) and water (340 mL) was heated at 50° C. till a clearsolution was formed. The mixture was allowed to cool to room temperatureand stood for 48 hours. The precipitate was collected by filtration,washed with ethanol and dried under reduced pressure to afford a saltwith L-camphorsulfonic acid (65.0 g). The salt was dissolved in water(500 mL) and 10% aqueous Na₂CO₃ (400 mL) was added. The mixture wasstirred for 1 hour and extracted with dichloromethane twice. Thecombined extracts were washed with brine (40 mL), dried over Na₂SO₄ andconcentrated under reduced pressure to provide(4S)-6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-amine(38.0 g).

Example 228a,b

N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-chloropyridine-2-carboxamide (352 mg, 0.821 mmol) was subjected tochromatography using supercritical CO₂ (supercritical CO₂/[EtOH with0.1% diethylamine]=60:40) on Chiralcel OD-H column (10×250 mm) elutingat a flow rate 10 mL/minute (40° C. column temperature). Afterconcentration of collected fractions of the first peak (retentiontime=5.23 minutes) at reduced pressure, recrystallization of the residuewith EtOH/water (1:1) provided a hydrate ofN-[(4R)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide(153 mg, 44%) as a crystal. After concentration of collected fractionsof the second peak (retention time=8.16 minutes) at reduced pressure,recrystallization of the residue with EtOH/water (1:1) provided ahydrate ofN-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide(152 mg) as a crystal.

Example 229a,b

N-[2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide(100 mg, 0.235 mmol) was chromatographed using supercritical CO₂(supercritical CO₂/EtOH=60:40) on Chiralcel OD-H column (10×250 mm)eluting at a flow rate 10 mL/minute (40° C. column temperature). Afterconcentration of collected fractions of the first peak (retentiontime=5.25 minutes) under reduced pressure, trituration of the residuewith EtOAc/hexane providedN-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide(35 mg). After concentration of collected fractions of the second peak(retention time=8.08 minutes) under reduced pressure, trituration of theresidue with EtOAc/hexane providedN-[(4R)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide(35 mg).

Example 230

To a solution of tert-butyl[(4′R)-6′-{[(5-methoxypyrazin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl]carbamate(13.75 g, 26.98 mmol) in chloroform (140 ml) was added trifluoroaceticacid (68 ml) in an ice-water bath, and the mixture was stirred at roomtemperature for 3 hours. The mixture was concentrated in vacuo, and theresidue was purified by silica gel column chromatography (pre-column:NH-silica gel; main column: neutral silica gel,chloroform/methanol=100:0-10:1). To the purified product was addedsaturated aqueous sodium hydrogen carbonate and extracted withchloroform. The organic layer were washed with brine, dried overanhydrous sodium sulfate, filtered, and the filtrate was evaporated invacuo. The residue was purified by silica gel column chromatography(pre-column: basic silica gel; main column: neutral silica gel,chloroform/methanol=100:0-10:1). The purified product was trituratedwith a mixture of hexane/ethyl acetate (4:1) (300 mL), and the mixturewas stirred at 60° C. for 1 hour and room temperature for 4 days. Theprecipitate was collected, washed with mixture of hexane/ethyl acetate(4:1) (200 mL), and dried in vacuo at 50° C. to affordN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide(8.38 g) as a crystal.

Example 231

To a solution ofN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide(800 mg, 1.86 mmol) in MeOH (10 mL) was added 4M solution of hydrogenchloride in ethyl acetate (0.5 mL, 2 mmol) and the mixture wasconcentrated in vacuo. The residue was triturated with EtOH (10 mL), andthe mixture was refluxed for 30 minutes and stirred at room temperatureovernight. The precipitate was collected, washed with EtOH (2 mL), anddried under reduced pressure at 70° C. overnight to affordN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamidehydrochloride (449 mg) as a crystal.

The compounds of Examples and Reference Examples shown in Tables belowwere prepared using the respective corresponding starting materials inthe same manner as the methods of Examples or Reference Examples above.The structures and the preparation methods are shown in [Table. 4]below, and the physicochemical data for the compounds of Examples orReference Examples are shown in [Table. 5] below.

TABLE 4 Ex Syn Structure RP1a RP1a, b

RP1b RP1a, b

RP2 RP6

RP3 RP6

RP4 RP6

RP5 RP6

RP6 RP6

RP7a RP1a, b

RP7b RP1a, b

RP8 RP6

RP9a RP1a, b

RP9b RP1a, b

RP10 RP6

RP11a RP1a, b

RP11b RP1a, b

RP12a RP1a, b

RP12b RP1a, b

RP13a RP1a, b

RP13b RP1a, b

RP14a RP1a, b

RP14b RP1a, b

RP15a RP1a, b

RP15b RP1a, b

RP16a RP1a, b

RP16b RP1a, b

RP17 RP6

RP18 RP6

RP19 RP19

RP20 RP19

RP21 RP19

RP22 RP19

RP23 RP19

RP24 RP19

RP25 RP19

RP26 RP19

27 E27

28 E27

RP29 E27

30 E27

RP31 RP31

RP32 RP31

RP33 RP39

RP34 RP39

RP35 RP39

RP36 RP39

RP37 RP39

RP38 RP39

RP39 RP39

RP40 RP40

RP41 RP41

RP42 RP41

RP43 RP43

RP44 RP43

RP45 RP43

RP46 RP46

RP47 RP46

RP48 RP48

RP49 RP49

RP50 RP49

RP51a RP51a, b

RP51b RP51a, b

RP52a RP52a, b

RP52b RP52a, b

RP53a RP53a, b

RP53b RP53a, b

RP54 RP54

RP55 RP55

RP56 RP61

RP57 RP61

RP58 RP61

RP59 RP61

RP60 RP61

RP61 RP61

RP62 RP61

RP63 RP61

RP64 RP61

RP65 RP65

RP66 RP65

RP67 RP65

RP68 RP19

RP69 RP75

RP70 RP75

RP71 RP75

RP72 RP75

RP73 RP75

RP74 RP75

RP75 RP75

RP76 RP75

RP77 RP75

RP78 RP75

RP79 RP75

RP80 RP75

RP81 RP75

RP82 RP75

RP83 RP75

RP84 RP75

RP85 RP75

RP86 RP75

RP87 RP75

RP88 RP75

RP89 RP75

RP90 RP75

RP91 RP75

RP92 RP75

RP93 RP75

RP94 RP75

RP95 RP75

RP96 RP75

RP97 RP75

RP98 RP75

RP99 RP75

RP100 RP75

RP101 RP75

RP102 RP75

RP103 RP75

RP104 RP75

RP105 RP75

RP106 RP75

RP107 RP75

RP108 RP108

RP109 RP108

RP110 RP108

RP111 RP108

RP112 RP108

RP113 RP108

RP114 RP114

RP115 RP115

RP116 RP116

RP117 RP116

RP118 RP116

RP119 RP136

RP120 RP136

RP121 RP136

RP122 RP136

RP123 RP136

RP124 RP136

RP125 RP136

RP126 RP136

RP127 RP136

RP128 RP136

RP129 RP136

RP130 RP136

RP131 RP136

RP132 RP136

RP133 RP136

RP134 RP136

RP135 RP136

RP136 RP136

RP137 RP136

RP138 RP136

RP139 RP146

RP140 RP146

RP141 RP146

RP142 RP146

RP143 RP146

RP144 RP146

RP145 RP146

RP146 RP146

RP147 RP146

RP148 RP146

RP149 RP146

RP150 RP146

RP151 RP146

RP152 RP146

RP153 RP146

RP154 RP146

RP155 RP146

RP156 RP146

RP157 RP146

RP158 RP146

RP159 RP146

RP160 RP146

RP161 RP146

RP162 RP146

RP163 RP146

RP164 RP146

RP165 RP146

RP166 RP146

RP167 RP146

RP168 RP146

RP169 RP174

RP170 RP174

RP171 RP174

RP172 RP174

RP173 RP174

RP174 RP174

RP175 RP174

RP176 RP174

RP177 RP174

RP178 RP174

RP179 RP174

RP180 RP174

RP181 RP174

RP182 RP174

RP183 RP174

RP184 RP174

RP185 RP174

RP186 RP174

RP187 RP174

RP188 RP6

RP189 E27

RP190 E27

RP191 E27

RP192 E27

RP193 E27

RP194 E27

RP195 E27

RP196 E27

RP197 E27

RP198 RP198

RP199 RP55

RP200 RP57

RP201 RP75

RP202 RP75

RP203 RP75

RP204 RP75

RP205 RP75

RP206 RP75

RP207 RP75

RP208 RP75

RP209 RP75

RP210 RP75

RP211 RP75

RP212 RP75

RP213 RP75

RP214 RP214

RP215 RP215

RP216 RP216

217 RP217

218 E218

219 E218

220 E218

221 E223

222 E223

223 E223

224 E224

RP225a RP225a, b

RP225b RP225a, b

RP226 RP226

227 E218

228a E228a, b

228b E228a, b

229a E229a, b

229b E229a, b

230 E230

231 E231

TABLE 5 Ex Data RP ESI+: 339, 341 1a less polar diastereomer RP ESI+:339, 341 1b polar diastereomer RP ESI+: 325, 327 2 RP ESI+: 365, 367 3RP ESI+: 397 4 RP ESI+: 365 5 RP ESI+: 353, 355 6 RP ESI+: 339, 341 7aRP ESI+: 339, 341 7b RP ESI+: 323, 325 8 RP ESI+: 353, 355 9a adiastereomer with higher Rf value on TLC (CHCl₃/EtOAc 9:1, NH silicagel)RP ESI+: 353, 355 9b a diastereomer with lower Rf value on TLC(CHCl₃/EtOAc 9:1, NH silicagel) RP ESI+: 379, 381 10 RP ESI+: 367, 36911a a diastereomer with higher Rf value on TLC (CHCl₃/EtOAc 9:1, NHsilicagel) RP ESI+: 367, 369 11b a diastereomer with lower Rf value onTLC (CHCl₃/EtOAc 9:1, NH silicagel) RP ESI+: 365, 367 12a a diastereomerwith higher Rf value on TLC (CHCl₃/EtOAc 9:1, NH silicagel) RP ESI+:365, 367 12b a diastereomer with lower Rf value on TLC (CHCl₃/EtOAc 9:1,NH silicagel) RP ESI+: 381, 383 13a a diastereomer with higher Rf valueon TLC (CHCl₃/EtOAc 9:1, NH silicagel) RP ESI+: 381, 383 13b adiastereomer with lower Rf value on TLC (CHCl₃/EtOAc 9:1, NH silicagel)RP ESI+: 421, 423 14a a diastereomer with higher Rf value on TLC(CHCl₃/EtOAc 9:1, NH silicagel) RP ESI+: 421, 423 14b a diastereomerwith lower Rf value on TLC (CHCl₃/EtOAc 9:1, NH silicagel) RP ESI+: 415,417 15a a diastereomer with higher Rf value on TLC (CHCl₃/EtOAc 9:1, NHsilicagel) RP ESI+: 415, 417 15b NMR-CDCl₃: 3.43 (1H, dd, J = 9.0, 14.7Hz), 3.53 (1H, dd, J = 3.1, 14.7 Hz), 3.99 (1H, d, J = 8.5 Hz),4.10-4.18 (2H, m), 4.47-4.55 (4H, m), 4.64 (1H, d, J = 6.7 Hz), 4.81(1H, d, J = 6.3 Hz), 6.63 (1H, d, J = 8.7 Hz), 7.21 (1H, dd, J = 2.4,8.7 Hz), 7.25-7.41 (6H, m) a diastereomer with lower Rf value on TLC(CHCl₃/EtOAc 9:1, NH silicagel) RP ESI+: 379, 381 16a a diastereomerwith higher Rf value on TLC (CHCl₃/EtOAc 9:1, NH silicagel) RP ESI+:379, 381 16b a diastereomer with lower Rf value on TLC (CHCl₃/EtOAc 9:1,NH silicagel) RP ESI+: 309, 311 17 RP ESI+: 327, 329 18 RP ESI+: 392 19NMR-DMSO-d₆: 3.37 (3H, s), 4.16-4.28 (5H, m), 4.37-4.39 (3H, m), 4.60(1H, d, J = 5.4 Hz), 4.71 (1H, d, J = 11.4 Hz), 6.40 (2H, s), 6.88 (1H,d, J = 8.4 Hz), 7.48 (1H, d, J = 2.3 Hz), 7.54 (1H, dd, J = 8.5, 2.4Hz), 8.02-8.03 (1H, m), 8.60 (1H, d, J = 1.9 Hz), 8.76 (1H, d, J = 2.3Hz) RP ESI+: 362 20 NMR-DMSO-d₆: 2.10 (3H, s), 4.17 (2H, m), 4.23-4.28(3H, m), 4.38 (1H, d, J = 6.4 Hz), 4.60 (1H, d, J = 5.3 Hz), 4.73 (1H,d, J = 11.4 Hz), 6.44 (2H, s), 6.88 (1H, d, J = 8.5 Hz), 7.51 (1H, dd, J= 5.3, 1.8 Hz), 7.55 (1H, d, J = 2.3 Hz), 7.58-7.61 (2H, m), 8.52 (1H,d, J = 5.2 Hz) RP ESI+: 363 21 NMR-DMSO-d₆: 2.16 (3H, s), 4.10-4.17 (2H,m), 4.23-4.30 (3H, m), 4.38 (1H, d, J = 6.4 Hz), 4.59 (1H, d, J = 5.4Hz), 4.75 (1H, d, J = 11.5 Hz), 6.46 (2H, s), 6.89 (1H, d, J = 8.6 Hz),7.97-8.00 (2H, m), 8.17 (1H, d, J = 2.3 Hz), 9.09 (1H, d, J = 1.2 Hz) RPESI+: 363 22 RP ESI+: 348 23 NMR-DMSO-d₆: 4.16-4.28 (5H, m), 4.38 (1H,d, J = 6.4 Hz), 4.49 (1H, s), 4.60 (1H, d, J = 5.4 Hz), 4.71 (1H, d, J =11.4 Hz), 6.40 (2H, s), 6.88 (1H, d, J = 8.5 Hz), 7.48 (1H, d, J = 2.3Hz), 7.53 (1H, dd, J = 8.4, 2.4 Hz), 8.04-8.05 (1H, m), 8.61 (1H, d, J =1.9 Hz), 8.78 (1H, d, J = 2.3 Hz) RP ESI+: 329 24 RP ESI+: 339 25 RPESI+: 369 26 NMR-DMSO-d₆: 3.85 (3H, s), 4.07 (1H, d, J = 8.8 Hz),4.13-4.16 (2H, m), 4.21-4.24 (2H, m), 4.34 (1H, d, J = 6.3 Hz),4.56-4.61 (2H, m), 6.31 (2H, s), 6.63 (1H, d, J = 8.8 Hz), 6.67 (1H, dd,J = 7.8, 5.0 Hz), 7.13 (1H, dd, J = 7.9, 1.4 Hz), 7.56 (1H, d, J = 2.6Hz), 7.61-7.64 (2H, m), 7.97 (1H, s)  27 ESI+: 429, 431 NMR-DMSO-d₆:1.18 (3H, s), 1.73 (3H, s), 4.13-4.19 (2H, m), 4.30-4.32 (2H, m), 4.36(1H, d, J = 6.8 Hz), 4.90 (1H, d, J = 5.6 Hz), 6.33 (2H, s), 6.66 (1H,d, J = 8.8 Hz), 7.57 (1H, dd, J = 8.8, 2.7 Hz), 7.70 (1H, d, J = 2.5Hz), 8.13 (1H, dd, J = 8.5, 0.7 Hz), 8,18 (1H, dd, J = 8.4, 2.4 Hz),8.75 (1H, dd, J = 2.3, 0.8 Hz), 10.52 (1H, s)  28 ESI+: 413 NMR-DMSO-d₆:1.18 (3H, s), 1.73 (3H, s), 4.13-4.19 (2H, m), 4.30-4.32 (2H, m), 4.36(1H, d, J = 6.8 Hz), 4.90 (1H, d, J = 5.6 Hz), 6.32 (2H, s), 6.66 (1H,d, J = 8.8 Hz), 7.55 (1H, dd, J = 8.8, 2.6 Hz), 7.70 (1H, d, J = 2.5Hz), 7.96 (1H, td, J = 8.7, 2.9 Hz), 8.20 (1H, dd, J = 8.7, 4.6 Hz),8.70 (1H, d, J = 2.8 Hz), 10.46 (1H, s) RP ESI+: 403, 405 29NMR-DMSO-d₆: 0.37-0.43 (3H, m), 0.82-0.85 (1H, m), 3.58 (1H, d, J = 11.6Hz), 4.09 (1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.2 Hz), 4.34-4.38 (1H,m), 6.18 (2H, s), 6.73-6.75 (1H, m), 7.55-7.57 (2H, m), 8.29 (1H, dd, J= 10.2, 1.9 Hz), 8.62-8.63 (1H, m), 10.47 (1H, s)  30 ESI+: 447, 449NMR-DMSO-d₆: 1.18 (3H, s), 1.73 (3H, s), 4.13-4.18 (2H, m), 4.31 (2H, d,J = 6.0 Hz), 4.36 (1H, d, J = 6.8 Hz), 4.89 (1H, d, J = 5.6 Hz), 6.34(2H, s), 6.65-6.68 (1H, m), 7.52-7.55 (2H, m), 8.29 (1H, dd, J = 10.2,2.0 Hz), 8.62-8.63 (1H, m), 10.51 (1H, s) RP ESI+: 369 31 NMR-DMSO-d₆:0.36-0.43 (3H, m), 0.82-0.88 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09(1H, d, J = 8.1 Hz), 4.30-4.37 (2H, m), 6.17 (2H, s), 6.73 (1H, d, J =8.8 Hz), 7.58 (1H, dd, J = 8.8, 2.6 Hz), 7.71 (1H, d, J = 2.6 Hz), 7.96(1H, td, J = 8.7, 2.9 Hz), 8.18-8.21 (1H, m), 8.70 (1H, d, J = 2.9 Hz),10.41 (1H, s) RP ESI+: 385, 387 32 NMR-DMSO-d₆: 0.36-0.44 (3H, m),0.82-0.88 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09 (1H, d, J = 8.1 Hz),4.31 (1H, d, J = 8.1 Hz), 4.35 (1H, dd, J = 11.5, 1.4 Hz), 6.17 (2H, s),6.73 (1H, d, J = 8.8 Hz), 7.59 (1H, dd, J = 8.8, 2.6 Hz), 7.71 (1H, d, J= 2.6 Hz), 8.12 (1H, dd, J = 8.4, 0.7 Hz), 8.18 (1H, dd, J = 8.5, 2.3Hz), 8.75 (1H, dd, J = 2.3, 0.7 Hz), 10.47 (1H, s) RP ESI+: 371 33 RPESI+: 376 34 RP ESI+: 319 35 RP ESI+: 325 36 RP ESI+: 338 37 RP ESI+:365 38 RP ESI+: 354 39 RP ESI+: 373, 375 40 RP ESI+: 341, 343 41 RPESI+: 369, 371 42 RP ESI+: 373, 375 43 NMR-DMSO-d₆: 4.05-4.36 (6H, m),4.57-4.65 (2H, m), 6.32 (2H, brs), 6.67-6.75 (2H, m), 7.39-7.42 (2H, m),7.70-7.72 (1H, m), 7.97-7.99 (1H, m), 8.26 (1H, s) RP ESI+: 413, 415 44NMR-DMSO-d₆: 1.82-1.88 (2H, m), 2.13-2.24 (2H, m), 2.32-2.43 (1H, m),2.92-2.98 (1H, m), 3.95-4.06 (1H, m), 4.16-4.21 (2H, m), 4.44-4.51 (2H,m), 4.62-4.66 (1H, m), 6.29 (2H, brs), 6.71-6.74 (2H, m), 7.31-7.38 (2H,m), 7.70-7.71 (1H, m), 7.98-7.99 (1H, m), 8.25 (1H, s) RP ESI+: 409 45NMR-DMSO-d₆: 1.80-1.88 (2H, m), 2.12-2.40 (3H, m), 2.89-2.97 (1H, m),3.76-3.94 (4H, m), 4.17-4.24 (2H, m), 4.49-4.64 (3H, m), 6.29 (2H, brs),6.67-6.72 (2H, m), 7.13 (1H, d, J = 7.6 Hz), 7.57-7.70 (3H, m), 8.00(1H, brs) RP ESI+: 343 46 RP APCI/ESI+: 371 47 RP ESI+: 428 48NMR-DMSO-d₆: 0.77-0.82 (2H, m), 0.90-0.96 (2H, m), 1.57-1.64 (1H, m),1.80-1.95 (2H, m), 2.12-2.34 (2H, m), 2.38-2.50 (1H, m), 2.93-3.04 (1H,m), 4.11-4.21 (3H, m), 4.49-4.53 (2H, m), 4.65 (1H, d, J = 5.5 Hz), 6.39(2H, brs), 6.91 (1H, d, J = 8.5 Hz), 7.27 (1H, d, J = 2.2 Hz), 7.50 (1H,dd, J = 8.5, 2.4 Hz), 7.86-7.87 (1H, m), 8.49 (1H, d, J = 1.9 Hz), 8.65(1H, d, J = 2.3 Hz) RP ESI+: 421 49 RP APCI/ESI+: 421 50 RP ESI+: 35351a retention time = 3.44 minutes RP ESI+: 353 51b NMR-DMSO-d₆: 1.22(3H, s), 1.77 (3H, s), 4.20 (1H, d, J = 8.9 Hz), 4.29-4.34 (3H, m), 4.39(1H, d, J = 6.9 Hz), 4.91 (1H, d, J = 5.6 Hz), 6.35 (2H, s), 6.85 (1H,d, J = 8.5 Hz), 7.35 (1H, d, J = 2.4 Hz), 7.54 (1H, dd, J = 8.4, 2.4Hz), 8.97 (2H, s), 9.14 (1H, s) retention time = 6.92 minutes RP ESI+:346 52a NMR-DMSO-d₆: 0.38-0.46 (3H, m), 0.81-0.88 (1H, m), 2.11 (3H, s),3.65 (1H, d, J = 11.7 Hz), 4.21 (1H, d, J = 8.1 Hz), 4.34 (1H, d, J =8.1 Hz), 4.41-4.45 (1H, m), 6.21 (2H, brs), 6.87 (1H, d, J = 8.5 Hz),7.37-7.41 (1H, m), 7.50-7.53 (1H, m), 7.91-7.92 (1H, t, J = 2.1 Hz),8.51 (1H, d, J = 2.1 Hz), 8.70 (1H, d, J = 2.1 Hz); retention time =3.62 minutes RP ESI+: 346 52b retention time = 6.27 minutes RP ESI+: 36253a retention time = 5.81 minutes RP ESI+: 362 53b NMR-DMSO-d₆: 2.11(3H, s), 4.15-4.21 (2H, m), 4.24-4.27 (3H, m), 4.38 (1H, d, J = 6.4 Hz),4.60 (1H, d, J = 5.3 Hz), 4.71 (1H, d, J = 11.4 Hz), 6.41 (2H, s), 6.87(1H, d, J = 8.5 Hz), 7.46 (1H, d, J = 2.3 Hz), 7.51 (1H, dd, J = 2.3,8.5 Hz), 7.92-7.94 (1H, m), 8.52 (1H, d, J = 1.9 Hz), 8.70 (1H, d, J =2.2 Hz); retention time = 9.25 minutes RP ESI+: 369 54 RP ESI+: 363 55RP ESI+: 362 56 NMR-DMSO-d₆: 2.12 (3H, s), 4.10-4.17 (2H, m), 4.24-4.27(3H, m), 4.38 (1H, d, J = 6.4 Hz), 4.59-4.61 (1H, m), 4.71 (1H, d, J =11.4 Hz), 6.43 (2H, s), 6.83 (1H, d, J = 8.6 Hz), 7.24 (1H, dd, J = 5.0,1.4 Hz), 7.756-7.762 (1H, m), 7.83 (1H, dd, J = 8.6, 2.4 Hz), 7.99 (1H,d, J = 2.3 Hz), 8.58 (1H, dd, J = 5.1, 0.8 Hz) RP ESI+: 348 57NMR-DMSO-d₆: 0.42 (3H, brs), 0.82 (3H, brs), 1.02-1.07 (2H, m),1.98-2.02 (1H, m), 3.65 (1H, d, J = 11.3 Hz), 4.20-4.22 (1H, m),4.32-4.36 (1H, m), 4.43 (1H, d, J = 11.3 Hz), 6.22 (2H, brs), 6.86 (1H,d, J = 8.4 Hz), 7.36 (1H, brs), 7.46-7.50 (2H, m), 8.32 (1H, s), 8.50(1H, s) RP ESI+: 406 58 NMR-DMSO-d₆: 0.41-0.46 (3H, m), 0.82-0.86 (1H,m), 3.65 (1H, d, J = 11.6 Hz), 4.21 (1H, d, J = 8.1 Hz), 4.34 (1H, d, J= 8.1 Hz), 4.44 (1H, d, J = 11.6 Hz), 4.92-4.99 (2H, m), 6.21 (2H, brs),6.89 (1H, d, J = 8.5 Hz), 7.41-7.43 (1H, m), 7.51-7.54 (1H, m),7.64-7.66 (1H, m), 8.34 (1H, d, J = 2.6 Hz), 8.43-8.46 (1H, m) RP ESI+:388 59 NMR-DMSO-d₆: 0.41-0.46 (3H, m), 0.82-0.88 (1H, m), 3.66 (1H, d, J= 11.7 Hz), 4.20 (1H, d, J = 8.16 Hz), 4.33 (1H, d, J = 8.16 Hz),4.42-4.54 (3H, m), 6.20 (2H, brs), 6.29-6.58 (1H, m), 6.88 (1H, d, J =8.5 Hz), 7.40-7.41 (1H, m), 7.51-7.53 (1H, m), 7.57-7.58 (1H, m),8.29-8.30 (1H, m), 8.40-8.41 (1H, m) RP ESI+: 374 60 NMR-DMSO-d₆:0.40-0.45 (3H, m), 0.82-0.85 (1H, m), 3.66 (1H, d, J = 11.6 Hz), 4.20(1H, d, J = 8.1 Hz), 4.34 (1H, d, J = 8.1 Hz), 4.44 (1H, d, J = 11.6Hz), 6.21 (2H, brs), 6.90 (1H, d, J = 8.4 Hz), 7.21-7.58 (3H, m),7.77-7.79 (1H, m), 8.42 (1H, d, J = 2.6 Hz), 8.66 (1H, d, J = 1.9 Hz) RPESI+: 347 61 NMR-DMSO-d₆: 0.41-0.46 (3H, m), 0.84-0.87 (1H, m), 3.62(1H, d, J = 11.7 Hz), 4.21 (1H, d, J = 8.1 Hz), 4.34 (1H, d, J = 8.1Hz), 4.42 (1H, d, J = 11.7 Hz), 6.20 (2H, brs), 6.48-6.50 (1H, m), 6.85(1H, d, J = 8.4 Hz), 7.36-7.37 (1H, m), 7.43-7.46 (1H, m), 7.48-7.49(1H, m), 8.03 (1H, d, J = 2.1 Hz), 8.37 (1H, d, J = 2.1 Hz), 11.6 (1H,brs) RP ESI+: 381, 383 62 RP ESI+: 336 63 RP ESI+: 438 64 RP ESI+: 38265 RP ESI+: 417 66 RP ESI+: 417 67 RP ESI+: 378 68 NMR-DMSO-d₆: 2.11(3H, s), 3.34-3.40 (2H, m), 4.23-4.28 (2H, m), 4.43 (1H, d, J = 11.3Hz), 4.54 (1H, d, J = 6.4 Hz), 4.63 (1H, d, J = 5.4 Hz), 4.74 (1H, d, J= 11.4 Hz), 6.87 (2H, s), 6.90 (1H, d, J = 8.5 Hz), 7.42 (1H, d, J = 2.3Hz), 7.53 (1H, dd, J = 8.6, 2.3 Hz), 7.87 (1H, t, J = 2.1 Hz), 8.52 (1H,d, J = 1.9 Hz), 8.66 (1H, d, J = 2.2 Hz) RP ESI+: 353 69 NMR-DMSO-d₆:1.22 (3H, s), 1.77 (3H, s), 4.20 (1H, d, J = 8.9 Hz), 4.29-4.35 (3H, m),4.39 (1H, d, J = 6.9 Hz), 4.91 (1H, d, J = 5.6 Hz), 6.35 (2H, s), 6.85(1H, d, J = 8.5 Hz), 7.35 (1H, d, J = 2.3 Hz), 7.54 (1H, dd, J = 8.6,2.3 Hz), 8.97 (2H, s), 9.14 (1H, s) RP ESI+: 382 70 RP ESI+: 382 71NMR-DMSO-d₆: 1.82-1.94 (2H, m), 2.14-2.33 (2H, m), 2.39-2.51 (1H, m),2.95-3.03 (1H, m), 4.06 (1H, d, J = 8.7 Hz), 4.17-4.20 (2H, m),4.49-4.53 (2H, m), 4.65 (1H, d, J = 5.4 Hz), 6.38 (2H, s), 6.93 (1H, d,J = 8.5 Hz), 7.25-7.26 (1H, m), 7.37-7.40 (1H, m), 7.42-7.46 (1H, m),7.96-8.01 (1H, m), 8.18-8.20 (1H, m) RP ESI+: 394 72 NMR-DMSO-d₆:1.81-1.94 (2H, m), 2.13-2.33 (2H, m), 2.38-2.51 (1H, m), 2.95-3.02 (1H,m), 3.89 (3H, s), 4.10 (1H, d, J = 8.6 Hz), 4.17-4.20 (2H, m), 4.50-4.54(2H, m), 4.65 (1H, d, J = 5.4 Hz), 6.39 (2H, s), 6.92 (1H, d, J = 8.4Hz), 7.28 (1H, d, J = 2.3 Hz), 7.43 (1H, dd, J = 2.6, 2.0 Hz), 7.50 (1H,dd, J = 8.4, 2.3 Hz), 8.25 (1H, d, J = 2.7 Hz), 8.32 (1H, d, J = 1.8 Hz)RP ESI+: 416 73 RP ESI+: 378 74 RP ESI+: 378 75 RP ESI+: 379 76 RP ESI+:356 77 RP ESI+: 339 78 RP ESI+: 356 79 RP ESI+: 339 80 RP ESI+: 368 81RP ESI+: 370 82 NMR-DMSO-d₆: 1.22 (3H, s), 1.77 (3H, s), 4.18 (1H, d, J= 8.9 Hz), 4.24 (1H, d, J = 8.9 Hz), 4.32-4.33 (2H, m), 4.39 (1H, d, J =6.9 Hz), 4.91 (1H, d, J = 5.6 Hz), 6.35 (2H, s), 6.82 (1H, d, J = 8.5Hz), 7.27-7.28 (1H, m), 7.34-7.38 (1H, m), 7.44 (1H, ddd, J = 7.4, 4.9,2.0 Hz), 7.99 (1H, ddd, J = 10.4, 7.5, 2.0 Hz), 8.18-8.20 (1H, m) RPESI+: 382 83 NMR-DMSO-d₆: 1.21 (3H, s), 1,77 (3H, s), 3.89 (3H, s), 4.18(1H, d, J = 8.8 Hz), 4.28-4.33 (3H, m), 4.40 (1H, d, J = 6.9 Hz), 4.91(1H, d, J = 5.6 Hz), 6.36 (2H, s), 6.81 (1H, d, J = 8.4 Hz), 7.30 (1H,d, J = 2.3 Hz), 7.44 (1H, dd, J = 2.7, 1.9 Hz), 7.47 (1H, dd, J = 8.5,2.3 Hz), 8.25 (1H, d, J = 2.7 Hz), 8.32 (1H, d, J = 1.8 Hz) RP ESI+: 35384 NMR-DMSO-d₆: 1.12 (3H, t, J = 7.2 Hz), 1.59-1.74 (1H, m), 1.86-1.99(1H, m), 4.20 (1H, d, J = 6.9 Hz), 4.38 (1H, d, J = 6.3 Hz), 4.42 (1H,d, J = 6.9 Hz), 4.45-4.52 (1H, m), 4.54-4.68 (3H, m), 6.22 (2H, bs),6.91 (1H, d, J = 8.4 Hz), 7.48 (1H, d, J = 2.3 Hz), 7.56 (1H, dd, J =2.3, 8.4 Hz), 9.02 (2H, s), 9.13 (1H, s) a compound prepared from Ex. 9aRP ESI+: 379 85 NMR-DMSO-d₆: 1.51-1.55 (1H, m), 1.60-1.77 (4H, m),1.87-1.96 (1H, m), 2.13-2.21 (1H, m), 2.49-2.56 (1H, m), 4.19-4.24 (2H,m), 4.27-4.29 (2H, m), 4.43 (1H, d, J = 6.8 Hz), 4.83 (1H, d, J = 5.3Hz), 6.36 (2H, s), 6.85 (1H, d, J = 8.5 Hz), 7.34 (1H, d, J = 2.3 Hz),7.53 (1H, dd, J = 8.5, 2.4 Hz), 8.97 (2H, s), 9.14 (1H, s) RP ESI+: 39086 NMR-DMSO-d₆: 1.11 (3H, t, J = 7.1 Hz), 1.57-1.72 (1H, m), 1.84-1.97(1H, m), 2.11 (3H, s), 4.20 (1H, d, J = 6.9 Hz), 4.37 (1H, d, J = 6.3Hz), 4.41 (1H, d, J = 6.9 Hz), 4.44-4.51 (1H, m), 4.52-4.66 (3H, m),6.23 (2H, bs), 6.87 (1H, d, J = 8.5 Hz), 7.41 (1H, d, J = 2.4 Hz), 7.50(1H, dd, J = 2.4, 8.5 Hz), 7.93-7.97 (1H, m), 8.52 (1H, d, J = 1.9 Hz),8.71 (1H, d, J = 2.3 Hz) a compound prepared from Reference Example 9aRP ESI+: 353 87 NMR-DMSO-d₆: 1.21 (3H, t, J = 7.3 Hz), 2.02-2.30 (2H,m), 3.95-4.13 (3H, m), 4.20-4.30 (1H, m), 4.33-4.45 (2H, m), 4.55 (1H,d, J = 5.5 Hz), 6.44 (2H, bs), 6.9 (1H, d, J = 8.5 Hz), 7.46 (1H, bs),7.51-7.61 (1H, m), 8.99 (2H, bs), 9.13 (1H, s) a compound prepared fromReference Example 9b RP ESI+: 365 88 NMR-DMSO-d₆: 0.39-0.51 (2H, m),0.67-0.81 (2H, m), 1.36-1.46 (1H, m), 3.80 (1H, d, J = 9.5 Hz), 4.15(1H, d, J = 7.0 Hz), 4.50 (1H, d, J = 6.4 Hz), 4.64-4.66 (2H, m), 4.81(2H, s), 6.21 (2H, s), 6.92 (1H, d, J = 8.4 Hz), 7.53-7.57 (2H, m), 9.04(2H, s), 9.13 (1H, s) a compound prepared from Reference Example 12a RPESI+: 365 89 a compound prepared from Reference Example 12b RP ESI+: 38190 NMR-DMSO-d₆: 1.01 (3H, d, J = 6.5 Hz), 1.07 (3H, d, J = 6.5 Hz),1.88-1.95 (1H, m), 2.02-2.15 (2H, m), 4.02 (1H, d, J = 8.9 Hz),4.11-4.14 (2H, m), 4.25 (1H, d, J = 5.5 Hz), 4.32 (1H, d, J = 6.5 Hz),4.41 (1H, d, J = 6.6 Hz), 4.58 (1H, d, J = 5.5 Hz), 6.44 (2H, s), 6.88(1H, d, J = 8.5 Hz), 7.45 (1H, d, J = 2.4 Hz), 7.54 (1H, dd, J = 8.5,2.4 Hz), 8.98 (2H, s), 9.13 (1H, s) a compound prepared from ReferenceExample 13b RP ESI+: 367 91 NMR-DMSO-d₆: 0.98 (3H, t, J = 7.2 Hz),1.42-1.58 (1H, m), 1.62-1.77 (2H, m), 1.78-1.90 (1H, m), 4.19 (1H, d, J= 6.8 Hz), 4.37 (1H, d, J = 6.4 Hz), 4.42 (1H, d, J = 6.8 Hz), 4.53-4.71(4H, m), 6.23 (2H, bs), 6.90 (1H, d, J = 8.4 Hz), 7.49 (1H, d, J = 2.3Hz), 7.55 (1H, dd, J = 2.3, 8.4 Hz), 9.02 (2H, s), 9.13 (1H, s) acompound prepared from Reference Example 11a RP ESI+: 404 92NMR-DMSO-d₆: 0.98 (3H, t, J = 7.2 Hz), 1.43-1.58 (1H, m), 1.61-1.75 (2H,m), 1.76-1.89 (1H, m), 2.11 (3H, s), 4.20 (1H, d, J = 7.0 Hz), 4.32-4.47(2H, m), 4.51-4.78 (4H, m), 6.24 (2H, bs), 6.86 (1H, d, J = 8.5 Hz),7.31-7.60 (2H, m), 7.97 (1H, bs), 8.52 (1H, d, J = 1.9 Hz), 8.72 (1H,bs) a compound prepared from Reference Example 11a RP ESI+: 381 93NMR-DMSO-d₆: 0.98 (3H, d, J = 6.6 Hz), 1.01 (3H, d, J = 6.7 Hz),1.62-1.68 (1H, m), 1.74-1.81 (1H, m), 1.93-2.03 (1H, m), 4.17 (1H, d, J= 7.0 Hz), 4.37 (1H, d, J = 6.5 Hz), 4.41 (1H, d, J = 7.0 Hz), 4.54 (1H,d, J = 6.5 Hz), 4.63-4.72 (3H, m), 6.23 (2H, s), 6.89 (1H, d, J = 8.5Hz), 7.49 (1H, d, J = 2.3 Hz), 7.55 (1H, dd, J = 8.5, 2.3 Hz), 9.02 (2H,s), 9.14 (1H, s) a compound prepared from Reference Example 13a RP ESI+:367 94 NMR-DMSO-d₆: 1.05 (3H, t, J = 7.4 Hz), 1.45-1.63 (1H, m),1.73-1.89 (1H, m), 2.00-2.21 (2H, m), 3.94-4.15 (3H, m), 4.23 (1H, d, J= 5.5 Hz), 4.37 (1H, d, J = 6.5 Hz), 4.41 (1H, d, J = 6.5 Hz), 4.56 (1H,d, J = 5.5 Hz), 6.44 (2H, bs), 6.89 (1H, d, J = 8.5 Hz), 7.46 (1H, bs),7.55 (1H, dd, J = 2.2, 8.5 Hz), 8.99 (2H, bs), 9.13 (1H, s) a compoundprepared from Reference Example 11b RP ESI+: 421 95 NMR-DMSO-d₆:1.90-2.07 (1H, m), 2.16-2.20 (1H, m), 2.52-2.69 (2H, m), 4.18 (1H, d, J= 7.2 Hz), 4.40 (1H, d, J = 6.8 Hz), 4.47 (1H, d, J = 7.2 Hz), 4.55 (1H,d, J = 6.8 Hz), 4.58-4.68 (1H, m), 4.73 (1H, d, J = 9.5 Hz), 4.75 (1H,d, J = 9.5 Hz), 6.21 (2H, bs), 6.96 (1H, d, J = 8.5 Hz), 7.53 (1H, d, J= 2.2 Hz), 7.58 (1H, dd, J = 2.2, 8.5 Hz), 9.04 (2H, s), 9.14 (1H, s) acompound prepared from Reference Example 14a RP ESI+: 458 96NMR-DMSO-d₆: 1.89-2.05 (1H, m), 2.11 (3H, s), 2.14-2.29 (1H, m),2.50-2.69 (2H, m), 4.18 (1H, d, J = 7.2 Hz), 4.39 (1H, d, J = 6.8 Hz),4.46 (1H, d, J = 7.2 Hz), 4.56 (1H, d, J = 6.8 Hz), 4.58-4.65 (1H, m),4.69 (1H, d, J = 9.2 Hz), 4.74 (1H, d, J = 9.2 Hz), 6.22 (2H, bs), 6.91(1H, d, J = 8.5 Hz), 7.45 (1H, d, J = 2.3 Hz), 7.52 (1H, dd, J = 2.3,8.5 Hz), 7.95-8.00 (1H, m), 8.52 (1H, d, J = 1.9 Hz), 8.72 (1H, d, J =2.3 Hz) a compound prepared from Reference Example 14a RP ESI+: 339 97 acompound prepared from Reference Example 1a RP ESI+: 339 98 a compoundprepared from Reference Example 1b RP ESI+: 432 99 RP ESI+: 309 100NMR-DMSO-d₆: 0.42-0.44 (3H, m), 0.81-0.83 (1H, m), 3.67 (1H, d, J = 11.6Hz), 4.21 (1H, d, J = 7.8 Hz), 4.35 (1H, d, J = 7.8 Hz), 4.44 (1H, d, J= 11.6 Hz), 6.21 (2H, brs), 6.91-6.93 (1H, d, J = 8.5 Hz), 7.45 (1H, s),7.55-7.58 (1H, m), 8.99 (2H, s), 9.12 (1H, s) RP ESI+: 323 101 RP ESI+:327 102 NMR-DMSO-d₆: 0.42-0.48 (3H, m), 0.81-0.86 (1H, m), 3.71 (1H, d,J = 11.7 Hz), 4.19 (1H, d, J = 8.2 Hz), 4.35 (1H, d, J = 8.2 Hz), 4.49(1H, dd, J = 1.6, 11.7 Hz), 6.20 (2H, brs), 6.83 (1H, d, J = 12.0 Hz),7.29 (1H, d, J = 9.0 Hz), 8.91 (2H, d, J = 1.4 Hz), 9.17 (1H, s) RPESI+: 326 103 NMR-DMSO-d₆: 0.39-0.46 (3H, m), 0.83-0.85 (1H, m), 3.65(1H, d, J = 10.9 Hz), 4.16 (1H, d, J = 8.0 Hz), 4.33 (1H, d, J = 8.0Hz), 4.44 (1H, d, J = 10.9 Hz), 6.20 (2H, brs), 6.89 (1H, d, J = 8.4Hz), 7.33-7.34 (1H, m), 7.37-7.40 (1H, m), 7.42-7.45 (1H, m), 7.97-8.02(1H, m), 8.17-8.18 (1H, m) RP ESI+: 360, 362 104 NMR-DMSO-d₆: 0.42-0.46(3H, m), 0.81-0.83 (1H, m), 3.67 (1H, d, J = 11.7 Hz), 4.16 (1H, d, J =8.2 Hz), 4.34 (1H, d, J = 8.2 Hz), 4.45 (1H, dd, J = 11.7, 1.6 Hz), 6.21(2H, brs), 6.89 (1H, d, J = 8.5 Hz), 7.37-7.38 (1H, m), 7.41-7.44 (1H,m), 8.16-8.19 (1H, m), 8.24-8.25 (1H, m) RP ESI+: 338 105 NMR-DMSO-d₆:0.41-0.45 (3H, m), 0.82-0.86 (1H, m), 3.65 (1H, d, J = 11.7 Hz), 3.89(3H, s), 4.20 (1H, d, J = 8.1 Hz), 4.32 (1H, d, J = 8.1 Hz), 4.41-4.44(1H, m), 6.21 (2H, brs), 6.87 (1H, d, J = 8.4 Hz), 7.37-7.38 (1H, m),7.44-7.45 (1H, m), 7.48-7.51 (1H, m), 8.24 (1H, d, J = 2.8 Hz), 8.34(1H, d, J = 1.9 Hz) RP ESI+: 352 106 NMR-DMSO-d₆: 0.42-0.44 (3H, m),0.82-0.84 (1H, m), 1.37 (3H, t, J = 7.0 Hz), 3.65 (1H, d, J = 11.7 Hz),4.15-4.20 (3H, m), 4.32-4.34 (1H, m), 4.41-4.44 (1H, m), 6.21 (2H, brs),6.87 (1H, d, J = 8.5 Hz), 7.38 (1H, s), 7.44 (1H, s), 7.48-7.50 (1H, m),8.22 (1H, d, J = 2.7 Hz), 8.33 (1H, d, J = 1.2 Hz) RP ESI+: 342, 344 107NMR-DMSO-d₆: 0.38-0.48 (3H, m), 0.81-0.84 (1H, m), 3.67 (2H, d, J = 11.9Hz), 4.20-4.25 (1H, m), 4.32-4.37 (1H, m), 4.45 (1H, d, J = 11.9 Hz),6.21 (2H, brs), 6.89 (1H, d, J = 8.2 Hz), 7.42-7.45 (1H, m), 7.55-7.59(1H, m), 8.09 (1H, brs), 8.55-8.56 (1H, m), 8.74 (1H, brs) RP ESI+: 362108 RP ESI+: 402 109 RP ESI+: 390 110 RP ESI+: 346 111 NMR-DMSO-d₆:0.41-0.45 (3H, m), 0.81-0.86 (1H, m), 2.11 (3H, s), 3.65 (1H, d, J =11.6 Hz), 4.21 (1H, d, J = 8.1 Hz), 4.33 (1H, d, J = 8.1 Hz), 4.43 (1H,d, J = 11.6 Hz), 6.21 (2H, brs), 6.87 (1H, d, J = 8.4 Hz), 7.39 (1H, d,J = 2.4 Hz), 7.50-7.53 (1H, m), 7.91 (1H, t, J = 2.1 Hz), 8.51 (1H, d, J= 2.1 Hz), 8.70 (1H, d, J = 2.1 Hz) RP ESI+: 364 112 NMR-DMSO-d₆:0.41-0.49 (3H, m), 0.81-0.88 (1H, m), 2.10 (3H, s), 3.69 (1H, d, J =11.8 Hz), 4.18-4.41 (2H, m), 4.48 (1H, d, J = 11.8 Hz), 6.22 (2H, brs),6.79 (1H, d, J = 11.8 Hz), 7.26 (1H, brs), 7.86 (1H, s), 8.56 (1H, d, J= 2.0 Hz), 8.59 (1H, s) RP ESI+: 360 113 NMR-DMSO-d₆: 1.57-1.62 (2H, m),1.68-1.76 (1H, m), 1.85-2.00 (2H, m), 2.11 (3H, s), 2.14-2.18 (1H, m),3.97-4.10 (3H, m), 4.37-4.43 (1H, m), 6.26 (2H, brs), 6.83-6.86 (1H, m),7.42 (1H, s), 7.48-7.50 (1H, m), 7.91 (1H, s), 8.50-8.51 (1H, m), 8.69(1H, s) RP ESI+: 339, 341 114 RP ESI+: 389 115 NMR-DMSO-d₆: 1.82-1.97(2H, m), 2.15-2.34 (2H, m), 2.44-2.52 (1H, m), 2.94-3.03 (1H, m), 4.10(1H, d, J = 8.7 Hz), 4.20 (1H, d, J = 5.5 Hz), 4.26 (1H, d, J = 8.7 Hz),4.48 (1H, d, J = 6.8 Hz), 4.53 (1H, d, J = 6.8 Hz), 4.66 (1H, d, J = 5.5Hz), 6.36 (2H, s), 6.99 (1H, d, J = 8.5 Hz), 7.28 (1H, d, J = 2.3 Hz),7.45 (1H, dd, J = 8.5, 2.3 Hz), 7.80 (1H, dd, J = 8.0, 4.7 Hz), 8.02(1H, dd, J = 8.0, 1.6 Hz), 8.72 (1H, dd, J = 4.7, 1.6 Hz) RP ESI+: 396116 RP ESI+: 379 117 RP ESI+: 414 118 RP ESI+: 382 119 RP ESI+: 389 120RP ESI+: 400 121 RP ESI+: 412 122 RP ESI+: 363 123 RP ESI+: 381 124 RPESI+: 381 125 RP ESI+: 393 126 RP ESI+: 377 127 RP ESI+: 388 128 RPESI+: 475 129 RP ESI+: 399 130 RP ESI+: 407 131 RP ESI+: 414 132 RPESI+: 425 133 RP ESI+: 406 134 RP ESI+: 367 135 RP ESI+: 411 136 RPESI+: 411 137 RP ESI+: 399 138 RP ESI+: 385, 387 139 RP ESI+: 391 140 RPESI+: 419 141 RP ESI+: 381 142 RP ESI+: 449 143 RP ESI+: 395 144 RPESI+: 411 145 RP ESI+: 403, 405 146 RP ESI+: 387 147 RP ESI+: 383 148 RPESI+: 370 149 RP ESI+: 387, 389 150 RP ESI+: 400, 402 151 RP ESI+: 420,422 152 RP ESI+: 404, 406 153 RP ESI+: 404, 406 154 RP ESI+: 416, 418155 RP ESI+: 386, 388 156 RP ESI+: 384 157 RP ESI+: 404, 406 158 RPESI+: 420, 422 159 RP ESI+: 377 160 RP ESI+: 400 161 RP ESI+: 416, 418162 RP ESI+: 386, 388 163 RP ESI+: 382 164 RP ESI+: 369 165 RP ESI+: 369166 RP ESI+: 343 167 RP ESI+: 391 168 RP ESI+: 385 169 RP ESI+: 394 170RP ESI+: 426 171 RP ESI+: 383 172 RP ESI+: 386, 388 173 RP ESI+: 390 174RP ESI+: 390 175 RP ESI+: 391 176 RP ESI+: 391 177 RP ESI+: 393 178 RPESI+: 402 179 RP ESI+: 402 180 RP ESI+: 345 181 RP ESI+: 395 182 RPESI+: 435 183 RP ESI+: 435 184 RP ESI+: 405 185 RP ESI+: 390 186 RPESI+: 417 187 RP ESI+: 359, 361 188 RP ESI+: 430, 432 189 NMR-DMSO-d₆:0.32-0.46 (3H, m), 0.78-0.90 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09(1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.36 (1H, dd, J = 1.3,11.6 Hz), 6.18 (2H, bs), 6.71-6.77 (1H, m), 7.58-7.65 (2H, m), 9.19 (2H,s), 10.60 (1H, s) 190 ESI+: 474, 476 NMR-DMSO-d₆: 1.18 (3H, s), 1.73(3H, s), 4.09-4.22 (2H, m), 4.25-4.42 (3H, m), 4.90 (1H, d, J = 5.6 Hz),6.33 (2H, bs), 6.63-6.71 (1H, m), 7.54-7.63 (2H, m), 9.19 (2H, s), 10.64(1H, s) RP ESI+: 390 191 NMR-DMSO-d₆: 0.34-0.45 (3H, m), 0.78-0.86 (1H,m), 2.52 (3H, s), 3.58 (1H, d, J = 11.6 Hz), 4.08 (1H, d, J = 8.1 Hz),4.31 (1H, d, J = 8.1 Hz), 4.36 (1H, dd, J = 1.5, 11.6 Hz), 6.18 (2H,bs), 6.71-6.76 (1H, m), 7.53-7.59 (2H, m), 8.34-8.38 (1H, m), 8.92-8.97(1H, m), 10.49 (1H, s) RP ESI+: 410, 412 192 NMR-DMSO-d₆: 0.35-0.45 (3H,m), 0.78-0.86 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09 (1H, d, J = 8.1Hz), 4.31 (1H, d, J = 8.1 Hz), 4.37 (1H, dd, J = 1.4, 11.6 Hz), 6.20(2H, bs), 6.76 (1H, d, J = 8.8 Hz), 7.45 (1H, d, J = 2.7 Hz), 7.55 (1H,dd, J = 2.7, 8.8 Hz), 8.77 (1H, d, J = 1.8 Hz), 9.07 (1H, d, J = 1.8Hz), 10.64 (1H, s) RP ESI+: 382 193 NMR-DMSO-d₆: 0.35-0.44 (3H, m),0.80-0.88 (1H, m), 3.58 (1H, d, J = 11.5 Hz), 4.01 (3H, s), 4.09 (1H, d,J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.34 (1H, br.d, J = 11.6 Hz),6.16 (2H, s), 6.72 (1H, d, J = 8.8 Hz), 7.56 (1H, dd, J = 2.7, 8.8 Hz),7.71 (1H, d, J = 2.6 Hz), 8.38 (1H, d, J = 1.3 Hz), 8.86 (1H, d, J = 1.3Hz), 10.30 (1H, s) RP ESI+: 399, 401 194 NMR-DMSO-d₆: 0.37-0.43 (3H, m),0.82-0.88 (1H, m), 2.54 (3H, s), 3.57 (1H, d, J = 11.7 Hz), 4.08 (1H, d,J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.34-4.37 (1H, m), 6.18 (2H, s),6.72 (1H, d, J = 8.7 Hz), 7.54-7.58 (2H, m), 7.98-7.99 (1H, m),8.536-8.543 (1H, m), 10.34 (1H, s) 195 ESI+: 443, 445 NMR-DMSO-d₆: 1.18(3H, s), 1.73 (3H, s), 2.53 (3H, s), 4.13-4.18 (2H, m), 4.30 (2H, d, J =6.1 Hz), 4.36 (1H, d, J = 6.8 Hz), 4.90 (1H, d, J = 5.6 Hz), 6.33 (2H,s), 6.64-6.66 (1H, m), 7.52-7.55 (2H, m), 7.99 (1H, dd, J = 2.4, 0.7Hz), 8.54-8.55 (1H, m), 10.38 (1H, s) RP ESI+: 376 196 NMR-DMSO-d₆:0.36-0.44 (3H, m), 0.82-0.85 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09(1H, d, J = 8.1 Hz), 4.30-4.37 (2H, m), 6.18 (2H, s), 6.74 (1H, d, J =8.8 Hz), 7.61 (1H, dd, J = 8.8, 2.7 Hz), 7.74 (1H, d, J = 2.6 Hz), 8.26(1H, dd, J = 8.2, 0.9 Hz), 8.56 (1H, dd, J = 8.2, 2.1 Hz), 9.16 (1H, dd,J = 2.1, 0.9 Hz), 10.64 (1H, s) 197 ESI+: 420 NMR-DMSO-d₆: 1.18 (3H, s),1.73 (3H, s), 4.13-4.19 (2H, m), 4.30-4.37 (3H, m), 4.90 (1H, d, J = 5.6Hz), 6.33 (2H, s), 6.67 (1H, d, J = 8.8 Hz), 7.59 (1H, dd, J = 8.8, 2.6Hz), 7.74 (1H, d, J = 2.5 Hz), 8.26 (1H, dd, J = 8.2, 0.9 Hz), 8.56 (1H,dd, J = 8.2, 2.1 Hz), 9.16-9.17 (1H, m), 10.69 (1H, s) RP ESI+: 375 198RP ESI+: 373 199 NMR-DMSO-d₆: 0.18-0.22 (1H, m), 0.39-0.46 (2H, m),0.61-0.63 (1H, m), 4.14 (1H, d, J = 11.5 Hz), 4.58 (1H, d, J = 11.5 Hz),6.47 (2H, brs), 6.82 (1H, s), 7.06 (1H, d, J = 8.7 Hz), 7.21-7.27 (2H,m), 7.42 (1H, s), 7.58 (1H, d, J = 8.7 Hz), 8.81 (2H, s), 9.05 (1H, s)RP ESI+: 415 200 NMR-DMSO-d₆: 2.89-2.95 (1H, m), 3.26-3.32 (1H, m), 4.30(1H, d, J = 7.0 Hz), 4.45 (1H, d, J = 6.3 Hz), 4.60-4.72 (4H, m),4.79-4.83 (1H, m), 6.28 (2H, brs), 6.82 (1H, d, J = 8.4 Hz), 7.23-7.29(1H, m), 7.32-7.37 (4H, m), 7.51 (1H, d, J = 2.3 Hz), 7.56 (1H, dd, J =2.3, 8.4 Hz), 9.02 (2H, s), 9.14 (1H, s) a compound prepared fromReference Example 15a RP ESI+: 452 201 NMR-DMSO-d₆: 2.11 (3H, s),2.87-2.94 (1H, m), 3.24-3.34 (1H, m), 4.31 (1H, d, J = 7.0 Hz), 4.44(1H, d, J = 6.3 Hz), 4.59-4.71 (4H, m), 4.79 (1H, dd, J = 1.9, 10.7 Hz),6.29 (2H, brs), 6.77 (1H, d, J = 8.5 Hz), 7.23-7.28 (1H, m), 7.32-7.37(4H, m), 7.44 (1H, d, J = 2.3 Hz), 7.51 (1H, dd, J = 2.3, 8.5 Hz),7.95-7.96 (1H, m), 8.52 (1H, d, J = 1.9 Hz), 8.71 (1H, d, J = 2.3 Hz) acompound prepared from Reference Example 15a RP ESI+: 415 202NMR-DMSO-d₆: 3.31-3.39 (1H, m), 3.57 (1H, brd, J = 13.5 Hz), 4.10-4.17(2H, m), 4.38-4.43 (2H, m), 4.48 (1H, d, J = 6.6 Hz), 4.53 (1H, d, J =6.6 Hz), 4.63 (1H, d, J = 5.7 Hz), 6.45 (2H, brs), 6.76 (1H, d, J = 8.4Hz), 7.24-7.28 (1H, m), 7.33-7.38 (2H, m), 7.46-7.53 (4H, m), 8.98 (2H,s), 9.13 (1H, s) a compound prepared from Reference Example 15b RP ESI+:452 203 NMR-DMSO-d₆: 2.10 (3H, s), 3.31-3.38 (1H, m), 3.56 (1H, brd, J =13.8 Hz), 4.10-4.15 (2H, m), 4.36-4.40 (1H, m), 4.42 (1H, d, J = 5.6Hz), 4.47 (1H, d, J = 6.6 Hz), 4.53 (1H, d, J = 6.6 Hz), 4.63 (1H, d, J= 5.6 Hz), 6.46 (2H, brs), 6.71 (1H, d, J = 8.4 Hz), 7.23-7.28 (1H, m),7.33-7.51 (6H, m), 7.89-7.91 (1H, m), 8.51 (1H, d, J = 1.9 Hz), 8.67(1H, d, J = 2.3 Hz) a compound prepared from Reference Example 15b RPESI+: 379 204 NMR-DMSO-d₆: 0.11-0.24 (2H, m), 0.43-0.58 (2H, m),1.00-1.08 (1H, m), 1.48-1.54 (1H, m), 1.82-1.90 (1H, m), 4.19 (1H, d, J= 6.9 Hz), 4.34 (1H, d, J = 6.3 Hz), 4.40 (1H, d, J = 6.9 Hz), 4.54-4.70(4H, m), 6.24 (2H, brs), 6.92 (1H, d, J = 8.5 Hz), 7.48 (1H, d, J = 2.3Hz), 7.57 (1H, dd, J = 2.3, 8.5 Hz), 9.02 (2H, s), 9.14 (1H, s) acompound prepared from Reference Example 16a RP ESI+: 416 205NMR-DMSO-d₆: 0.11-0.24 (2H, m), 0.43-0.58 (2H, m), 0.98-1.08 (1H, m),1.46-1.52 (1H, m), 1.80-1.87 (1H, m), 2.11 (3H, s), 4.19 (1H, d, J = 6.9Hz), 4.33 (1H, d, J = 6.2 Hz), 4.39 (1H, d, J = 6.9 Hz), 4.51 (1H, d, J= 9.0 Hz), 4.58-4.63 (2H, m), 4.65-4.69 (1H, m), 6.25 (2H, brs), 6.87(1H, d, J = 8.5 Hz), 7.41 (1H, d, J = 2.3 Hz), 7.51 (1H, dd, J = 2.3,8.5 Hz), 7.95 (1H, dd, J = 1.9, 2.2 Hz), 8.52 (1H, d, J = 1.9 Hz), 8.71(1H, d, J = 2.2 Hz) a compound prepared from Reference Example 16a RPESI+: 359 206 RP ESI+: 379 207 NMR-DMSO-d₆: 0.24-0.33 (2H, m), 0.44-0.50(1H, m), 0.56-0.62 (1H, m), 1.09-1.19 (1H, m), 1.69-1.75 (1H, m),2.30-2.37 (1H, m), 4.05-4.12 (2H, m), 4.15-4.18 (1H, m), 4.25 (1H, d, J= 5.5 Hz), 4.30 (1H, d, J = 6.6 Hz), 4.39 (1H, d, J = 6.6 Hz), 4.55 (1H,d, J = 5.5 Hz), 6.43 (2H, brs), 6.91 (1H, d, J = 8.4 Hz), 7.46 (1H, d, J= 2.4 Hz), 7.57 (1H, dd, J = 2.4, 8.4 Hz), 8.99 (2H, s), 9.13 (1H, s) acompound prepared from Reference Example 16b RP ESI+: 359 208 RP ESI+:364 209 NMR-DMSO-d₆: 1.80-1.95 (2H, m), 2.13-2.24 (1H, m), 2.24-2.34(1H, m), 2.37-2.48 (1H, m), 2.94-3.04 (1H, m), 4.07-4.15 (1H, m),4.15-4.23 (2H, m), 4.47-4.54 (2H, m), 4.66 (1H, d, J = 5.2 Hz), 6.40(2H, brs), 6.92 (1H, d, J = 8.4 Hz), 7.28 (1H, s), 7.44-7.50 (2H, m),7.90 (1H, d, J = 7.6 Hz), 8.52 (1H, d, J = 4.8 Hz), 8.73 (1H, s) RPESI+: 414 210 RP ESI+: 414 211 RP ESI+: 432 212 RP ESI+: 361 213 RPESI+: 323, 325 214 RP ESI+: 323 215 NMR-DMSO-d₆: 0.20-0.24 (1H, m),0.42-0.54 (2H, m), 0.95-0.98 (1H, m), 3.51 (1H, d, J = 11.6 Hz), 3.73(1H, d, J = 11.4 Hz), 3.84 (1H, d, J = 11.4 Hz), 3.97 (1H, d, J = 15.6Hz), 4.13 (1H, d, J = 15.6 Hz), 4.61 (1H, dd, J = 1.8, 11.6 Hz), 5.69(2H, brs), 6.89 (1H, d, J = 8.4 Hz), 7.31 (1H, d, J = 2.4 Hz), 7.53 (1H,dd, J = 2.4, 8.4 Hz), 8.99 (2H, s), 9.12 (1H, s) RP ESI+: 359, 361 216RP ESI+: 337, 339 217 218 ESI+: 429, 431 NMR-DMSO-d₆: 1.18 (3H, s), 1.73(3H, s), 4.13-4.19 (2H, m), 4.30-4.32 (2H, m), 4.36 (1H, d, J = 6.8 Hz),4.90 (1H, d, J = 5.6 Hz), 6.33 (2H, s), 6.66 (1H, d, J = 8.8 Hz), 7.57(1H, dd, J = 8.8, 2.6 Hz), 7.70 (1H, d, J = 2.6 Hz), 8.13 (1H, dd, J =8.5, 0.8 Hz), 8.18 (1H, dd, J = 8.5, 2.3 Hz), 8.75 (1H, dd, J = 2.4, 0.7Hz), 10.52 (1H, s) Melting point: 165° C. (differential scanningcalorimetry onset temperature, Heating rate: 10° C./minute, under N₂flow of 50 mL/minute) Crystals having characteristic peaks of powderX-ray diffraction shown at angles 20 (°) of about 5.7, 9.6, 11.4, 12.3,13.7, 15.7, 15.9 and 25.0. This is the same compound as Ex. 228b. 219ESI+: 473, 475 NMR-DMSO-d₆: 1.18 (3H, s), 1.73 (3H, s), 4.13-4.19 (2H,m), 4.30-4.32 (2H, m), 4.36 (1H, d, J = 6.7 Hz), 4.90 (1H, d, J = 5.6Hz), 6.33 (2H, s), 6.66 (1H, d, J = 8.8 Hz), 7.57 (1H, dd, J = 8.8, 2.6Hz), 7.70 (1H, d, J = 2.5 Hz), 8.05 (1H, dd, J = 8.4, 0.3 Hz), 8.29-8.32(1H, m), 8.82-8.85 (1H, m), 10.52 (1H, s) 220 ESI+: 426 NMR-DMSO-d₆:1.18 (3H, s), 1.73 (3H, s), 4.01 (3H, s), 4.14 (1H, d, J = 8.8 Hz), 4.17(1H, d, J = 8.8 Hz), 4.28-4.33 (2H, m), 4.35 (1H, d, J = 6.8 Hz), 4.90(1H, d, J = 5.6 Hz), 6.32 (2H, s), 6.65 (1H, d, J = 8.8 Hz), 7.53 (1H,dd, J = 2.6, 8.8 Hz), 7.72 (1H, d, J = 2.5 Hz), 8.38 (1H, d, J = 1.3Hz), 8.87 (1H, d, J = 1.4 Hz), 10.35 (1H, s) This is the same compoundas Ex. 229b. 221 ESI+: 413 NMR-CDCl₃: 0.42-0.55 (2H, m), 0.74-0.82 (1H,m), 1.18 (3H, s), 1.23-1.30 (1H, m), 1.33 (3H, s), 4.45 (br s), 4.46(1H, d, J = 8.0 Hz), 4.57 (1H, d, J = 8.0 Hz), 6.81 (1H, d, J = 8.8 Hz),7.45 (1H, d, J = 2.6 Hz), 7.60 (1H, dd, J = 2.6, 8.8 Hz), 7.84 (1H, dd,J = 2.4, 8.4 Hz), 8.20-8.24 (1H, m), 8.50-8.54 (1H, m), 9.69 (1H, bs)222 ESI+: 397 NMR-CDCl₃: 0.41-0.55 (2H, m), 0.73-0.82 (1H, m), 1.18 (3H,s), 1.23-1.30 (1H, m), 1.33 (3H, s), 4.45 (br s), 4.46 (1H, d, J = 8.0Hz), 4.57 (1H, d, J = 8.0 Hz), 6.81 (1H, d, J = 8.8 Hz), 7.44-7.47 (1H,m), 7.52-7.63 (2H, m), 8.28-8.33 (1H, m), 8.41 (1H, d, J = 2.8 Hz), 9.67(1H, bs) 223 ESI+: 410 NMR-DMSO-d₆: 0.23-0.32 (1H, m), 0.49-0.57 (1H,m), 0.59-0.68 (1H, m), 1.07-1.16 (1H, m), 1.11 (3H, s), 1.27 (3H, s),4.01 (3H, s), 4.26 (1H, d, J = 8.0 Hz), 4.37 (1H, d, J = 8.0 Hz), 6.09(2H, bs), 6.66 (1H, d, J = 8.7 Hz), 7.53 (1H, dd, J = 2.6, 8.7 Hz), 7.61(1H, d, J = 2.6 Hz), 8.38 (1H, d, J = 1.3 Hz), 8.86 (1H, d, J = 1.3 Hz),10.29 (1H, s) This is the same compound as Ex. 230. 224 ESI+: 430NMR-DMSO-d₆: 0.24-0.32 (1H, m), 0.50-0.58 (1H, m), 0.60-0.68 (1H, m),1.08-1.16 (1H, m), 1.12 (3H, s), 1.27 (3H, s), 4.27 (1H, d, J = 8.0 Hz),4.38 (1H, d, J = 8.0 Hz), 6.11(2H, bs), 6.69 (1H, d, J = 8.7 Hz), 7.24(1H, t, J = 53.9 Hz), 7.59 (1H, dd, J = 2.6, 8.7 Hz), 7.65 (1H, d, J =2.6 Hz), 9.06 (1H, s), 9.34-9.37 (1H, m), 10.71(1H, s) This is the freeform of Ex. 231. RP ESI+: 337, 339 225a HPLC retention time: 4.5 minutes(CHIRALCEL OD-RH, MeCN: 20 mM aqueous KH₂PO₄ = 80:20; flow rate 1.0ml/minute, detected by 254 nm UV absorption, 2nd peak of the enantiomerpair) RP ESI+: 337, 339 225b HPLC retention time: 2.4 minutes (CHIRALCELOD-RH, MeCN: 20 mM aqueous KH₂PO₄ = 80:20; flow rate 1.0 ml/minute,detected by 254 nm UV absorption, 1st peak of the enantiomer pair) RPESI+: 353, 355 226 227 ESI+: 426 228a ESI+: 429, 431 Supercritical fluidchromatography retention time: 5.23 minutes: (eluent: supercriticalCO₂/(EtOH with 0.1% diethylamine) = 60:40; Column: CHIRALCEL OD-H column(10 × 250 mm); flow rate 10 mL/minute; column temperature: 40° C., 1stpeak of the enantiomer pair) 228b ESI+: 429, 431 Supercritical fluidchromatography retention time: 8.16 minutes (eluent: supercritical CO₂/(EtOH with 0.1% diethylamine) = 60:40; Column: CHIRALCEL OD-H column (10× 250 mm); flow rate 10 mL/minute; column temperature: 40° C., 2nd peakof the enantiomer pair) This is the same compound as Ex. 218. 229a ESI+:426 Supercritical fluid chromatography retention time: 5.94 minutes(eluent: supercritical CO₂/EtOH = 60:40; Column: CHIRALCEL OD-H column(4.6 × 250 mm); flow rate 3 mL/minute; column temperature: 40° C., 2ndpeak of the enantiomer pair) 229b ESI+: 426 Supercritical fluidchromatography retention time: 3.58 minutes (eluent: supercritical CO₂/EtOH = 60:40; Column: CHIRALCEL OD-H column (4.6 × 250 mm); flow rate 3mL/minute; column temperature: 40° C., 1st peak of the enantiomer pair)This is the same compound as Ex. 220. 230 ESI+: 410 NMR-DMSO-d₆:0.23-0.32 (1H, m), 0.49-0.57 (1H, m), 0.59-0.68 (1H, m), 1.07-1.16 (1H,m), 1.11 (3H, s), 1.27 (3H, s), 4.01 (3H, s), 4.26 (1H, d, J = 8.0 Hz),4.37 (1H, d, J = 8.0 Hz), 6.10 (2H, bs), 6.66 (1H, d, J = 8.7 Hz), 7.53(1H, dd, J = 2.6, 8.7 Hz), 7.61 (1H, d, J = 2.6 Hz), 8.38 (1H, d, J =1.3 Hz), 8.86 (1H, d, J = 1.3 Hz), 10.29 (1H, s) Melting point: 191° C.(differential scanning calorimetry onset temperature, Heating rate: 10°C./minute, under N₂ flow of 50 mL/minute) Crystals having characteristicpeaks of powder X-ray diffraction shown at angles 20 (°) of about 5.0,7.9, 8.0, 8.8, 12.6, 15.2, 16.3, 17.7 and 20.2. This is the samecompound as Ex. 223. 231 ESI+: 430 NMR-DMSO-d₆: 0.54-0.66 (1H, m),0.76-0.92 (2H, m), 0.94-1.05 (1H, m), 1.18 (3H, s), 1.30 (3H, s), 5.07(2H, s), 6.87 (1H, d, J = 8.9 Hz), 7.26 (1H, t, J = 53.9 Hz), 7.90 (1H,dd, J = 2.4, 8.9 Hz), 7.98 (1H, d, J = 2.4 Hz), 9.02-9.21 (2H, m),9.37-9.41 (1H, m), 9.57 (1H, bs), 10.55 (1H, s), 10.98 (1H, s) Meltingpoint: 254° C. (differential scanning calorimetry onset temperature,Heating rate: 10° C./minute, under N₂ flow of 50 mL/minute) Crystalshaving characteristic peaks of powder X-ray diffraction shown at angles20 (°) of about 4.8, 6.5, 8.4, 12.8, 16.0, 17.4, 23.4, 26.6 and 27.6.This is the hydrochloride of Ex. 224.

The compounds shown in Tables below can be prepared using tert-butyl(6′-aminodispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-2″-yl)carbamateor tert-butyl(6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-2-yl)carbamateas a starting material in the same manner as the methods of PreparationExamples 70 and Reference Examples 19 or Examples 27 above. Thestructures, and the preparation methods, for the compounds are shown in[Table. 6] below.

TABLE 6 No. Structure 1st_step 2nd_step P1

R70 E27 P2

R70 E27 P3

R70 E27 P4

R70 E27 P5

R70 E27 P6

R70 E27 P7

R70 E27 P8

R70 E27 P9

R70 E27 P10

R70 E27 P11

R70 E27 P12

R70 E27 P13

R70 E27 P14

R70 E27 P15

R70 E27 P16

R70 E27 P17

R70 E27 P18

R70 E27 P19

R70 E27 P20

R70 E27 P21

R70 E27 P22

R70 E27 P23

R70 E27 P24

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The teachings of all references cited in this specification areincorporated herein in their entirety.

INDUSTRIAL APPLICABILITY

The compounds of the formula (I) or a salt thereof have a beta-secretaseinhibitory activity, and therefore can be used as an agent forpreventing or treating diseases or conditions associated with and/ormediated by β-secretase activity, hydrolysis of a β-secretase site of aβ-amyloid precursor protein, and/or β-amyloid protein accumulation, suchas Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease,especially, Alzheimer's disease, or the like.

The invention claimed is:
 1. A compound of the formula (I) or a saltthereof:

wherein A¹ is O; A² is —C(R^(A21)R^(A22))—; and R^(A21) and R^(A22) areH; B is

X is lower alkyl; Y is lower alkyl; R¹ and R⁴ are H; R³ is H or halogen;and R² is —N(H)—C(O)-(hetero ring group), wherein said hetero ring groupis unsubstituted or substituted with one or more substituents selectedfrom the group consisting of halogen, lower alkyl which is unsubstitutedor substituted with halogen, —CN, and —O-(lower alkyl).
 2. The compound,or a salt thereof, of claim 1, wherein R² is selected from the groupconsisting of —N(H)—C(O)-(pyridyl), wherein said pyridyl isunsubstituted or substituted with one or more substituents selected fromthe group consisting of halogen, lower alkyl, and —CN,—N(H)—C(O)-(pyrazinyl), wherein said pyrazinyl is unsubstituted orsubstituted with —O-(lower alkyl) or lower alkyl which is unsubstitutedor substituted with halogen, and —N(H)—C(O)-(pyrimidinyl), wherein saidpyrimidinyl is unsubstituted or substituted with halogen.
 3. Thecompound, or a salt thereof, of claim 2, wherein B is


4. The compound, or a salt thereof, of claim 3, wherein X is methyl; andY is methyl.
 5. The compound, or a salt thereof, of claim 4, wherein R³is H; and R² is —N(H)—C(O)-(pyridyl), wherein said pyridyl isunsubstituted or substituted with halogen.
 6. The compound, or a saltthereof, of claim 2, wherein B is


7. The compound, or a salt thereof, of claim 6, wherein X is methyl; andY is methyl.
 8. The compound, or a salt thereof, of claim 7, wherein R³is H; and R² is —N(H)—C(O)-(pyrazinyl), wherein said pyrazinyl isunsubstituted or substituted with —O-(lower alkyl) or lower alkyl whichis unsubstituted or substituted with halogen.
 9. The compound, or a saltthereof, of claim 1, which is selected from the group consisting ofN-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-chloropyridine-2-carboxamide,N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-fluoropyridine-2-carboxamide,N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-chloro-3-fluoropyridine-2-carboxamide,N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-bromopyrimidine-2-carboxamide,N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-chloro-3-methylpyridine-2-carboxamide,N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-cyanopyridine-2-carboxamide,N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl)-5-methoxypyrazine-2-carboxamide,N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide,N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide,N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide,N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide,N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-bromopyridine-2-carboxamide,N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-chloropyridine-2-carboxamide,andN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-fluoropyridine-2-carboxamide.10. The compound, or a salt thereof according to claim 1, wherein saidcompound is selected from the group consisting ofN-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide,N-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide,andN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide.11. The compound, or a salt thereof, of claim 1, wherein said compoundisN-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide.12. The compound, or a salt thereof, of claim 1, wherein said compoundisN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide.13. The compound, or a salt thereof, of claim 1, wherein said compoundisN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide.14. A pharmaceutical composition comprising a compound, or apharmaceutically acceptable salt thereof, according to claim 1 and apharmaceutically acceptable carrier.
 15. The pharmaceutical compositionof claim 14, wherein the compound isN—[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3″-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide,or a pharmaceutically acceptable salt thereof.
 16. The pharmaceuticalcomposition of claim 14, wherein the compound isN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide,or a pharmaceutically acceptable salt thereof.
 17. The pharmaceuticalcomposition of claim 14, wherein the compound isN-[(4′R)-2″-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4″-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide,or a pharmaceutically acceptable salt thereof.